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Phenotypic plasticity, the ability to produce multiple phenotypes from a single genotype, represents an excellent model with which to examine the relationship between gene expression and phenotypes. Analyses of the molecular foundations of phenotypic plasticity are challenging, however, especially in the case of complex social phenotypes. Here we apply a machine learning approach to tackle this challenge by analyzing individual-level gene expression profiles of Polistes dominula paper wasps following the loss of a queen. We find that caste-associated gene expression profiles respond strongly to queen loss, and that this change is partly explained by attributes such as age but occurs even in individuals that appear phenotypically unaffected. These results demonstrate that large changes in gene expression may occur in the absence of outwardly detectable phenotypic changes, resulting here in a socially mediated de-differentiation of individuals at the transcriptomic level but not at the levels of ovarian development or behavior., Connecting genotypes to complex social behaviour is challenging. Taylor et al. use machine learning to show a strong response of caste-associated gene expression to queen loss, wherein individual wasp’s expression profiles become intermediate between queen and worker states, even in the absence of behavioural changes.

The gut microbiota is recognized as having major impact in health and disease. Sample storage is an important aspect to obtain reliable results. Mostly recommended is immediate freezing, however, this is not always feasible. Faecal occult blood test (FOBT) papers are an appealing solution in such situations, and most studies find these to be applicable, showing no major changes within 7 days storage at room temperature (RT). As fieldwork often requires RT storage for longer periods, evaluation of this is warranted. We performed 16S rRNA gene sequencing of 19 paired faecal samples immediately frozen or kept five weeks and five months at RT on FOBT papers. Alpha-diversity evaluation revealed no effect of FOBT storage, and evaluation of beta-diversity showed that host explained 65% of community variation, while storage method explained 5%. Evaluation of community dispersion and the Firmicutes/Bacteroidetes ratio revealed a larger effect of storage time for fresh-frozen samples. Single taxa evaluation (order-to-genus level) showed significant alterations of four (of 37) genera after five weeks and five genera after five months. When comparing the two timepoints, alterations were only detectable for fresh-frozen samples. Our findings reveal that long term storage on FOBT papers is an applicable approach for microbiota research.

Relatively little is known about the processes shaping population structure in cooperatively breeding insect species, despite the long-hypothesized importance of population structure in shaping patterns of cooperative breeding. Polistes paper wasps are primitively eusocial insects, with a cooperative breeding system in which females often found nests in cooperative associations. Prior mark-recapture studies of Polistes have documented extreme female philopatry, although genetic studies frequently fail to detect the strong population structure expected for highly philopatric species. Together these findings have led to lack of consensus on the degree of dispersal and population structure in these species. This study assessed population structure of female Polistes fuscatus wasps at three scales: within a single site, throughout Central New York, and across the Northeastern United States. Patterns of spatial genetic clustering and isolation by distance were observed in nuclear and mitochondrial genomes at the continental scale. Remarkably, population structure was evident even at fine spatial scales within a single collection site. However, P. fuscatus had low levels of genetic differentiation across long distances. These results suggest that P. fuscatus wasps may employ multiple dispersal strategies, including extreme natal philopatry as well as longer-distance dispersal. We observed greater genetic differentiation in mitochondrial genes than in the nuclear genome, indicative of increased dispersal distances in males. Our findings support the hypothesis that limited female dispersal contributes toward population structure in paper wasps.

We investigated associative learning of food odors by the European paper wasp Polistes dominula Christ (Hymenoptera: Vespidae) because of consistent low rates of attraction to food materials in laboratory assays. We hypothesized that wasps in nature exhibit nonspecific food-finding behavior until locating a suitable food, and then respond more strongly and specifically to odors associated with that food reward. Female P. dominula workers exhibited higher rates of attraction in a flight tunnel to piped odors of fermented fruit purees following previous experience with that puree, compared to wasps with no prior experience with the fermented fruits. Attraction behavior included upwind-oriented flight and casting within the odor plume, indicative of chemoanemotaxis. Synthetic chemicals representative of volatiles P. dominula may encounter in nature while foraging was also tested. Similar increases in attraction responses occurred following feeding experience with a sugar solution that included either 3-methyl-1-butanol or pear ester, but not eugenol. These experimental results support the hypothesis of associative learning of food odors in P. dominula. We discuss the ecological relevance of our results and suggest an alternative approach to trap paper wasps in pest situations utilizing learned chemical attractants.

Social interactions have large effects on individual physiology and fitness. In the immediate sense, social stimuli are often highly salient and engaging. Over longer time scales, competitive interactions often lead to distinct social ranks and differences in physiology and behavior. Understanding how initial responses lead to longer-term effects of social interactions requires examining the changes in responses over time. Here we examined the effects of social interactions on transcriptomic signatures at two times, at the end of a 45-minute interaction and 4 hours later, in female Polistes fuscatus paper wasp foundresses. Female P. fuscatus have variable facial patterns that are used for visual individual recognition, so we separately examined the transcriptional dynamics in the optic lobe and the non-visual brain. Results demonstrate much stronger transcriptional responses to social interactions in the non-visual brain compared to the optic lobe. Differentially regulated genes in response to social interactions are enriched for memory-related transcripts. Comparisons between winners and losers of the encounters revealed similar overall transcriptional profiles at the end of an interaction, which significantly diverged over the course of 4 hours, with losers showing changes in expression levels of genes associated with aggression and reproduction in paper wasps. On nests, subordinate foundresses are less aggressive, do more foraging and lay fewer eggs compared to dominant foundresses and we find losers shift expression of many genes in the non-visual brain, including vitellogenin, related to aggression, worker behavior, and reproduction within hours of losing an encounter. These results highlight the early neurogenomic changes that likely contribute to behavioral and physiological effects of social status changes in a social insect., Author summary Aggressive interactions often create inequalities–some individuals win while others lose. Winning versus losing can lead to large physiological differences between individuals, including different neurogenomic profiles between winners and losers. How this information about contest outcome leads to distinct neurogenomic profiles is poorly understood. Here we examine gene expression in response to aggressive social encounters in paper wasps, which naturally form dominance hierarchies on their nests in the wild. Shortly following encounters winners and losers have similar expression profiles, likely because similar mechanisms are engaged by social experiences. Four hours later, we find divergent neurogenomic profiles between winners and losers, with losers showing larger shifts in expression compared to winners. Many of the most dynamically expressed genes have been previously associated with dominance and caste differences in paper wasps showing how a single interaction can engage many of the same genomic networks that are involved in mediating more dramatic differences in queen-worker behavioral differences are also involved in responses shortly following social interactions.

International audience; The incidence of arbovirus infections has increased dramatically in recent decades, affecting hundreds of millions of people each year. The Togaviridae family includes the chikungunya virus (CHIKV), which is typically transmitted by Aedes mosquitoes and causes a wide range of symptoms from flu-like fever to severe arthralgia. Although conventional diagnostic tests can provide early diagnosis of CHIKV infections, access to these tests is often limited in developing countries. Consequently, there is an urgent need to develop efficient, affordable, simple, rapid, and robust diagnostic tools that can be used in point-of-care settings. Early diagnosis is crucial to improve patient management and to reduce the risk of complications. A glass-fiber laser-cut microfluidic device (paper-based analytical device [PAD]) was designed and evaluated in a proof of principle context, for the analysis of 30 µL of patient serum. Biological raw materials used for the functionalization of the PAD were first screened by MAC-ELISA (IgM capture enzyme-linked immunosorbent assay) for CHIKV Immunoglobulin M (IgM) capture and then evaluated on the PAD using various human samples. Compared with viral lysate traditionally used for chikungunya (CHIK) serology, CHIKV pseudo-particles (PPs) have proven to be powerful antigens for specific IgM capture. The PAD was able to detect CHIKV IgM in human sera in less than 10 minutes. Results obtained in patient sera showed a sensitivity of 70.6% and a specificity of around 98%. The PAD showed few cross-reactions with other tropical viral diseases. The PAD could help health workers in the early diagnosis of tropical diseases such as CHIK, which require specific management protocols in at-risk populations.

Vermicomposting of paper mill sludge (PMS) and tea waste (TW) using cow dung (CD) in five different combinations was carried out using Eisenia fetida. The aim of this study was to manage the waste disposal problem of PMS using the environment-friendly technology of vermiconversion. The changes in physico-chemical parameters were observed at 30-day intervals up to 90 days. The final pH was within 6.09–6.95 among all units. The TN, TP and TK contents increased 0.30–0.87, 0.53–3.23, 0.33–0.63 times, respectively in all mixtures after vermicomposting with increase in EC and ash Content. Maximum reduction in Total Organic Carbon (23.91%) was observed in treatment with highest PMS content, attributed to earthworm activity. Reduction in C: N ratio (38.63%–54.05%) was significantly observed in all the treatments. It was finally inferred that the paper mill sludge and tea waste in combination with cow dung can be successfully biotransformed into useful manure employing earthworms.

Detection of Leishmania in cutaneous lesions is possible by visualization of amastigotes. Detection of Leishmania DNA by PCR presents greater sensitivity, and PCR has been used to diagnose cutaneous leishmaniosis in humans using noninvasive clinical specimens.Study I: to determine if Leishmania DNA could be efficiently extracted and amplified from archived Diff-QuikSamples from cutaneous lesions of 54 dogs.Study I: Leishmania-qPCR was performed on 19 glass slides (from nine dogs) with cytologically visible amastigotes. Fifteen slides with no visible amastigotes, obtained from 12 dogs seronegative for Leishmania by ELISA, served as controls. Study II: Leishmania-qPCR was performed on glass slides and FPI from cutaneous lesions compatible with clinical leishmaniosis in 33 dogs.Study I: all slides with visible amastigotes had positive qPCR, whereas all control slides yielded negative results. Study II: of 13 dogs definitively diagnosed with clinical leishmaniosis, eight had visible amastigotes on cytology, whereas Leishmania-qPCR was positive on 11 glass slides and 13 FPI. Leishmaniosis was ruled out by standard methods in 20 dogs, four of which yielded positive qPCR on FPI and/or glass slides.Leishmania-DNA can be detected efficiently by qPCR from cutaneous cytological specimens and FPI to diagnose Leishmania infection in dogs with cutaneous lesions suggestive of CanL.Détection de Leishmania dans les lésions cutanées est possible par visualisation des amastigotes. La détection de l’ADN de Leishmania par PCR présente une meilleure sensibilité et la PCR a été utilisée pour diagnostiquer la leishmaniose cutanée de l'homme à l'aide de techniques cliniques non invasives.Etude I : déterminer si l’ADN de Leishmania peut être efficacement extraite et amplifiée de lames colorées au Diff-QuikEchantillons de lésions cutanées de 54 chiens. MATÉRIEL ET MÉTHODE: Etude I : la PCRq a été réalisée sur 19 lames (de 9 chiens) avec des amastigotes visibles à la cytologie. Quinze lames sans amastigote visible, obtenu de 12 chiens séronégatifs pour Leishmania par ELISA, ont servi de contrôle. Etude II : Une PCRq Leishmania a été réalisée sur des lames et FPI de lésions cutanées compatibles avec une leishmaniose clinique chez 33 chiens. RÉSULTATS: Etude I : toutes les lames avec amastigotes visibles avaient une PCRq positive tandis que les lames contrôles étaient négatives. Etude II : sur les 13 chiens diagnostiqués définitivement comme leishmaniose clinique, huit avaient des amastigotes visibles à la cytologie tandis que la PCRq Leishmania était positive sur 11 des lames et 13 FPI. LA leishmaniose a été exclue par méthodes standard chez 20 chiens, dont quatre présentaient une PCRq positive sur FPI et/ou lames.L’ADN de Leishmania peut être détectée efficacement par PCRq de cytologies cutanées et FPI pour le diagnostic de leishmaniose chez les chiens présentant des lésions cutanées évocatrices de CanL.INTRODUCCIÓN: la visualización de amastigotes permite la detección de Leishmania en lesiones cutáneas. La detección del DNA de Leishmania por PCR es más sensible, y la PCR se ha utilizado para diagnosticar la leishmaniasis cutánea en humanos utilizando muestras clínicas no invasivas. OBJETIVOS: Estudio I: determinar si el DNA de Leishmania se podría extraer y amplificar de manera eficiente a partir de preparaciones teñidas con Diff-Quik® archivadas de muestras citológicas de lesiones cutáneas caninas. Estudio II: evaluar el valor diagnóstico de una PCR cuantitativa (qPCR) de Leishmania en muestras citológicas teñidas y en impresiones de papel de filtro (FPI) obtenidas de lesiones cutáneas sugestivas de leishmaniasis canina (CanL). ANIMALES: muestras de lesiones cutáneas de 54 perros. MÉTODOS Y MATERIALES: Estudio I: se realizó qPCR para Leishmania en 19 portaobjetos de vidrio (de nueve perros) con amastigotes citológicamente visibles. Quince preparaciones sin amastigotes visibles, obtenidas de 12 perros seronegativos para Leishmania por ELISA, sirvieron como controles. Estudio II: se realizó qPCR para Leishmania en portaobjetos de vidrio y FPI de lesiones cutáneas compatibles con leishmaniasis clínica en 33 perros. RESULTADOS: Estudio I: todas las preparaciones con amastigotes visibles tuvieron qPCR positiva, mientras que todas las preparaciones de control dieron resultados negativos. Estudio II: de 13 perros diagnosticados definitivamente con leishmaniasis clínica, ocho tuvieron amastigotes visibles en la citología, mientras que qPCR para Leishmania fue positiva en 11 portaobjetos de vidrio y 13 FPI. La leishmaniasis se descartó mediante métodos estándar en 20 perros, cuatro de los cuales produjeron qPCR positiva en FPI y/o portaobjetos de vidrio. CONCLUSIONES E IMPORTANCIA CLÍNICA: la qPCR para DNA de Leishmania puede realizarse de forma efectiva a partir de muestras citológicas cutáneas y FPI para diagnosticar la infección por Leishmania en perros con lesiones cutáneas sugestivas de CanL.Leishmanien können in Hautveränderungen durch eine Sichtbarmachung der Amastigoten gefunden werde. Die Feststellung von Leishmania DNA mittels PCR bedeutet eine größere Sensibilität. Außerdem wurde die PCR verwendet, um kutane Leishmaniose bei Menschen mittels nichtinvasiver klinischer Proben zu diagnostizieren.Studie I: festzustellen, ob ausreichend Leishmania DNA aus archivierten Diff-Quick® gefärbten zytologischen Präparaten aus kutanen Veränderungen extrahiert und amplifiziert werden konnte. Studie II: eine Evaluierung der diagnostischen Leistung einer quantitativen Leishmania (q) PCR aus gefärbten Zytologiepräparaten sowie aus Abdrücken auf Filterpapier (FPI), die aus Hautveränderungen stammten, die verdächtig für eine canine Leishmaniose (CanL) waren.Es wurden Proben von Hautveränderungen von 54 Hunden verwendet.Studie I: Es wurde ein Leishmania-qPCR von 19 Objektträgern (von neun Hunden) mit zytologisch sichtbaren Amastigoten durchgeführt. Fünfzehn Objektträger ohne sichtbare Amastigoten, die von 12 Hunden stammten, die mittels ELISA seronegativ auf Leishmania waren, dienten als Kontrollen. Studie II: Ein Leishmania-qPCR wurde von Objektträgern und FPI kutaner Läsionen von 33 Hunden, die mit klinischer Leishmaniose kompatibel waren, durchgeführt.Studie I: alle Objektträger mit sichtbaren Amastigoten zeigten eine positive qPCR, während die Kontrollen ein negatives Ergebnis lieferten. Studie II: von den 13 Hunden, die definitiv mit einer Leishmaniose diagnostiziert worden waren, zeigten acht sichtbare Amastigoten in der Zytologie, während der Leishmania-PCR von 11 Objektträgern und 13 FPI positiv war. Eine Leishmaniose war bei 20 Hunden mit Standardmethoden ausgeschlossen worden, von denen vier eine positive qPCR von FPI und/oder Objektträgern lieferten.Leishmania-DNA kann effektiv mittels qPCR aus kutanen zytologischen Hautproben und aus FPI festgestellt werden, um eine Leishmania Infektion bei Hunden mit Hautveränderungen, die auf CanL hinweisen zu diagnostizieren.背景: 皮膚病変におけるリーシュマニアの検出は、無鞭毛期虫体の視覚化によって可能となる。 PCRによるリーシュマニアDNAの検出はより高い感度を示し、PCRは非侵襲性臨床標本を用いて人の皮膚リーシュマニア症を診断するために使用されてきた。 目的: 研究I:研究Iの目的は、犬の皮膚病変から得られた細胞学的標本のアーカイブされたDiff-Quik®染色スライドからLeishmania DNAを効率的に抽出および増幅できるかどうかを決定することである。研究II:研究IIの目的は、染色した細胞学的標本および犬リーシュマニア症(CanL)を示唆する皮膚病変から得られたfilter paper impressions (FPI)に対するリーシュマニア定量的(q)PCRの診断性能を評価することである。 被験動物: 54頭の犬の皮膚病変からのサンプル。 材料と方法: 研究I:Leishmania-qPCRを、細胞学的に観察可能な無鞭毛期虫体を含む19枚のスライドガラス(9頭の犬から)によって実施した。 ELISAによってリーシュマニアに対して血清陰性の12頭の犬から得られた、目に見える無鞭毛期虫体のない15枚のスライドを対照として用いた。研究II:33頭の犬において、リーシュマニアqPCRを、臨床的にリーシュマニア症と適合性のある皮膚病変からのスライドガラスおよびFPIに対して実施した。 結果: 研究I:目に見える無鞭毛期虫体を含む全てのスライドがqPCR陽性であったのに対し、全対照スライドは陰性結果をもたらした。研究II:臨床リーシュマニア症と確定診断された13頭の犬のうち、8頭が細胞診で目に見える斑点があったのに対し、Leishmania-qPCRは11枚のスライドガラスと13枚のFPIで陽性を示した。リーシュマニア症は、20頭の犬において標準的な方法で除外され、そのうちの4頭はFPIおよび/またはスライドガラス上でqPCR陽性を示した。 結論と臨床的重要性: リーシュマニアDNAは、CanLを示唆する皮膚病変を有する犬におけるリーシュマニア感染を診断するために、皮膚細胞診標本およびFPIからqPCRによって効率的に検出することができる。.背景: 通过发现无鞭毛体可以检测皮肤病变中的利什曼原虫。 PCR检测利什曼原虫DNA具有更高的灵敏度,并且无创临床样本的PCR检测已被用于诊断人类皮肤利什曼病。 目的: 研究I:确认是否能从已被Diff-Quik®染色的犬皮肤病变细胞学载玻片中,有效提取和扩增利什曼原虫DNA。研究II:染色细胞学和滤纸印迹(FPI)均采自疑似犬利什曼病(CanL)的皮肤病变,评估利什曼原虫定量(q)PCR方法对这些样本的诊断效果。 动物: 54只犬的皮肤病变样本。 方法和材料: 研究I:细胞学检查可见无鞭毛体的19张载玻片(来自9只犬),对其进行利什曼原虫-qPCR检测;从 12只犬身上获得的15张载玻片样本作为对照,细胞学检查未见无鞭毛体,经ELISA检测利什曼原虫血清阴性。研究II:33只症状符合利什曼病的犬,用利什曼原虫-qPCR法检测其皮肤病变载玻片和FPI。 结果: 研究I:所有可见无鞭毛体的载玻片,其qPCR检测均为阳性,而所有对照载玻片的检测结果均为阴性。 研究II: 13只确诊利什曼病的患犬中,8只细胞学上可见无鞭毛体,而利什曼原虫-qPCR检测中11张载玻片和13份FPI呈阳性。用标准方法排除利什曼病的20只犬中,经qPCR检测FPI和/或载玻片,有4只呈阳性。 结论和临床价值: 用qPCR技术检测皮肤细胞学和FPI中的利什曼原虫DNA,可有效诊断皮肤病变疑似CanL的利什曼原虫感染。.A detecção de Leishmania em lesões cutâneas é possível pela visualização de amastigotas. A detecção de DNA de Leishmania por PCR possui maior sensibilidade, e a PCR tem sido utilizada para diagnosticar leishmaniose cutânea em humanos utilizando espécimens clínicos não invasivos.Estudo I: determinar se o DNA de Leishmania pode ser eficientemente extraído e amplificado de lâminas coradas em Diff-QuikAmostras cutâneas de lesões de 54 cães. MÉTODOS E MATERIAIS: Estudo I: Leishmania-qPCR foi realizado em 19 lâminas de vidro (de nove cães) com amastigotas citologicamente visíveis. Quinze lâminas sem amastigotas visíveis, obtidas de 12 cães soronegativos para Leishmania por ELISA serviram como controle. Estudo II: Leishmania-qPCR foi realizado em lâminas de vidro e em FPI de lesões cutâneas compatíveis com leishmaniose clínica em 33 cães.Estudo I: todas as amostras com amastigotas visíveis apresentaram qPCR positivo, enquanto todas as lâminas controle tiveram resultados negativos. Estudo II: dos 13 cães diagnosticados definitivamente com leishmaniose clínica, oito apresentaram amastigotas visíveis na citologia, enquanto Leishmania-qPCR foi positivo em 11 lâminas de vidro e 13 FPI. Descartou-se a leishmaniose pelos métodos convencionais em 20 cães, dos quais quatro apresentaram qPCR positiva na FPI e/ou nas lâminas de vidro. CONCLUSÕES E IMPORTÂNCIA CLÍNICA: O DNA de Leishmania pode ser eficientemente detectado por qPCR de espécimens citológicos cutâneos e FPI para diagnosticar uma infecção por Leishmania em cães com lesões cutâneas sugestivas de CanL.

Cooperation among kin is common in animal societies. Kin groups may form by individuals directly discriminating relatives based on kin recognition cues, or form passively through natal philopatry and limited dispersal. We describe the genetic landscape for a primitively eusocial wasp, Polistes dominula, and ask whether individuals choose cooperative partners that are nearby and/or that are genetic relatives. Firstly, we genotyped an entire sub-population of 1361 wasps and found genetic structuring on an extremely fine scale: the probability of finding genetic relatives decreases exponentially within just a few meters of an individual’s nest. At the same time, however, we found a lack of genetic structuring between natural nest aggregations within the population. Secondly, in a separate dataset where ~2000 wasps were genotyped, we show that wasps forced experimentally to make a new nest choice tended to choose new nests near to their original nests, and that these nests tended to contain some full sisters. However, a significant fraction of wasps chose nests that did not contain sisters, despite sisters being present in nearby nests. Although we cannot rule out a role for direct kin recognition or natal nest-mate recognition, our data suggest that kin groups may form via a philopatric rule-of-thumb, whereby wasps simply select groups and nesting sites that are nearby. The result is that most subordinate helpers obtain indirect fitness benefits by breeding cooperatively.

High mountains are harsh environments in which colder temperatures and higher levels of UV-B radiation are common. These abiotic conditions strongly affect animals’ biology, often constraining their survival and reproduction. As a result, adaptations to live in such habitats are expected to evolve. Body color is thought to be adaptive to the environment that animals experience. Tegument melanization improves heat gain and provides photoprotection. Therefore, at high elevation, ectotherms are expected to be darker (well-melanized). We test this prediction in the paper wasp Agelaia pallipes (Hymenoptera: Vespidae), a species distributed across an elevational gradient in the Colombian Andes. We used Malaise traps and sampled a total of 146 wasps along nine elevations, ranging from 2,600–3,380 m above sea level. Standard digital photography was used to measure the body luminance and colour patterning in different body parts of dry-preserved specimens. There was striking variation in body luminance (darker and lighter), color patterning (patched, smoothed, homogeneous) and surface texture (shiny and matte), but the kind and degree of variation depended on the body part examined. Wasps from higher elevations had darker thoraces, confirming our prediction. Besides, at high elevation, the frequency of wasps with a matte rather than a shiny face strongly increased. Overall, our findings support the thermal melanism hypothesis and suggest that intraspecific color variation might be an adaptation to the environment of paper wasps.

Pulp and paper mill effluents (PPMEs) have been shown to increase gonad size, cause early maturation, and disrupt hormone functions in native and non-native Chilean fish. In this study, we assessed reproductive (plasma vitellogenin; VTG, gonad development) and metabolic (ethoxyresorufin-. O-deethylase activity; EROD) end points, relative liver size (LSI) and condition factor (K) of juvenile female and male rainbow trout exposed to effluents. Unlike previous studies, which have focus either on the specific effects of effluent on fish in laboratory exposures or biotic population statuses downstream of discharge sites, we simultaneously assessed the impacts of PPMES on trout using two approaches: (1) laboratory exposures of tertiary treated PPME produced from processing Eucalyptus globulus or Pinus radiata; and (2) in situ bioassay downstream of the combined discharge of the same pulp mill. Despite an increase in the average gonadosomatic index (GSI) in exposed fish, no statistical differences in gonad size between exposed and unexposed individuals was detected. However, both female and male fish exposed to effluents showed significantly higher concentrations of plasma VTG, so more in fish exposed to Eucalyptus-based effluent when compared to Pinus PPME. In addition, male fish showed intersex characteristics in all exposure assays (Eucaliptus and Pinus) and, despite the low concentration of effluent in the river (

Vermitechnology is gaining recognition as an environmental friendly waste management strategy. Its successful implementation requires that the key operational parameters like earthworm stocking density be established for each target waste/waste mixture. One target waste mixture in South Africa is waste paper mixed with cow dung and rock phosphate (RP) for P enrichment. This study sought to establish optimal Eisenia fetida stocking density for maximum P release and rapid bioconversion of RP enriched cow dung-paper waste mixtures. E. fetida stocking densities of 0, 7.5, 12.5, 17.5 and 22.5 g-worms kg(-1) dry weight of cow dung-waste paper mixtures were evaluated. The stocking density of 12.5 g-worms kg(-1) resulted in the highest earthworm growth rate and humification of the RP enriched waste mixture as reflected by a C:N ratio of12 and a humic acid/fulvic acid ratio of1.9 in final vermicomposts. A germination test revealed that the resultant vermicompost had no inhibitory effect on the germination of tomato, carrot, and radish. Extractable P increased with stocking density up to 22.5 g-worm kg(-1) feedstock suggesting that for maximum P release from RP enriched wastes a high stocking density should be considered.

Exposure to early adverse experiences induces persistent changes in physiological, emotional and behavioural functions predisposing the individual to an enhanced vulnerability to develop different disorders during lifespan. The adverse outcomes depend upon the timing of the stressful experiences, and in this contest, adolescence represents a key sensitive period for brain development. Among the biological systems involved, gut microbiota has recently been proposed to act on the interplay between the stress response, brain functions and immune system, through the gut-brain axis communication. In the current study we aimed to evaluate, in a preclinical model, changes over time in the microbiota community structure in physiological condition and in response to stress during adolescence. We also aimed to correlate the microbiota composition to the inflammatory status in brain. We used the preclinical model of social deprivation in rats during adolescence, based on the lack of all social contacts, for four weeks after weaning, followed by re-socialization until adulthood. We collected fecal samples at different post-natal days to investigate the short- and long-lasting effects of social isolation on gut microbiota composition and we collected brain areas (dorsal and ventral hippocampus) samples at killing to measure a panel of inflammatory and microglia activation markers. 16 S metataxonomic sequencing analysis revealed that microbial changes were influenced by age in both isolated and controls rats, regardless of sex, whereas social isolation impacted the microbial composition in a sex-dependent manner. A multivariate analysis showed that social isolation induced short-term gut microbiota alterations in females but not in males. We also identified several stress-related genera associated with social isolation condition. In brain areas we found a specific inflammatory pattern, in dorsal and ventral hippocampus, that significantly correlated with gut microbiota composition. Overall, in this study we reported a novel sex-specific association between gut microbiota composition and inflammatory response related to social isolation paradigm during adolescence, suggesting that stressful experiences during this sensitive period could have a long-lasting impact on the development of different biological systems that could in turn influence the vulnerability to develop mental disorders later in life.

The prevailing paradigm for social wasp endocrinology is that of juvenile hormone (JH) functioning pleiotropically in potential and actual queens, where it fuels dominance behaviors, stimulates ovarian growth and/or affects the production of status-linked cuticular compounds. In colonies with annual cycles (e.g., temperate-zone species), female adults produced at the end of the summer (called gynes) are physiologically primed to hibernate. Despite the absence of egg-laying in the pre-overwintering phase, gynes engage in dominance interactions that may affect reproductive potential following hibernation. JH levels have long been inferred to be low in gynes but this has never been tested. In what is the first study to measure JH in gyne-containing colonies of a temperate paper wasp, and the first to incorporate hormone assays in Belonogaster, our results show that the JH titer positively correlates with gyne-specific traits (including oocyte length and a low frequency of foraging trips) in B. longitarsus, a South African paper wasp. Measures of dominance correlated with oocyte length, but not all dominant females possessed activated ovaries. The cuticular hydrocarbon profiles of gynes and workers were distinct, with oocyte length and JH titer showing a positive association with longer-chain methyl-branched alkanes. Nonetheless, evidence for a role of JH in dominance was inconclusive. Finally, the range of JH titers among gynes, and the positive association of JH titers with ovarian status and prospective fertility signals, makes it unlikely that the gyne phenotype is maintained by low JH levels.

Little research has been conducted to examine the influence of various methods of providing nest materials—such as dispersing them, providing them as single units, or clustering them—on the behavior and welfare of group-housed mice. In this study, 6 wk-old C57BL/6NCrl mice were housed 3 per cage and randomized into 1 of 3 nest-material groups: 1) one facial tissue per cage (control; female mice, 3 cages; male mice, 3 cages); 2) an 8-g 'puck' of compressed nesting material and a facial tissue (females, 3 cages; males, 3 cages); or 3) 8 g of dispersed paper strips and a facial tissue (females, 3 cages; males, 3 cages). Mouse behavior (agonistic, stereotypic, nesting), physical examination data, and nest scores were evaluated over 16 d. The results showed that mice in the puck and control groups spent more time manipulating nest materials after cage changes than did mice in the paper-strip group. Average nest scores were highest in the paper-strip group compared with controls and puck cages. Female cages with pucks showed no barbering, whereas all other female mice cages demonstrated barbering. Overall, nest pucks may provide a time-consuming activity for mice and may help protect female C57BL/6 mice from barbering. However, more research is needed to replicate and expand these study results.

Several eusocial wasps are prominent invaders to remote islands. The paper wasp Polistes chinensis antennalis is native to East Asia, was introduced to New Zealand in 1979 and has expanded its distribution there. This provides an excellent opportunity to examine the impacts of an initial bottleneck and subsequent expansion on genetic structure. We analysed and compared the genetic population structures of the native (Japan and South Korea) and invasive New Zealand populations. Although 94% of individuals had shared haplotypes detected across both populations, the remaining 6% had private haplotypes identified in only one of the three countries. The genetic variation at microsatellite loci was lower in New Zealand than in native countries, and the genetic structure in New Zealand was clearly distinct from that in its native range. Higher frequencies of diploid-male- and triploid-female-producing colonies were detected in New Zealand than in the native countries, showing the reduction in genetic variation via a genetic bottleneck. At least two independent introductions were suggested, and the putative source regions for New Zealand were assigned as Kanto (central island) and Kyushu (south island) in Japan. Serial founder events following the initial introduction were also indicated. The estimated dispersal distance between mother and daughter in New Zealand was twice that in Japan. Thus, the introduction history of P. chinensis antennalis in New Zealand is probably the result of at least two independent introductions, passing through a bottleneck during introduction, followed by population expansion from the point of introduction.

The present clinical study aimed to investigate if there are differences in microbiological outcomes dependent on the subgingival biofilm collection method. Subgingival biofilm samples were collected from the four deepest pockets (>5 mm) of 17 patients with aggressive periodontitis (AgP) and 33 patients with chronic periodontitis (CP), first by paper point and thereafter by curette. Samples obtained with the same method were pooled together from each patient and forwarded for molecular microbiological analysis by a commercially available assay (IAI Pado Test 4.5) that estimates total bacterial load and levels of Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola and Aggregatibacter actinomycetemcomitans. Data analysis included frequency of detection, quantification and correlation of detection levels between the two sampling methods. P. gingivalis, T. forsythia and T. denticola were detected in >90% of the samples, and their detection levels exhibited a strong correlation between sampling methods. The detection consistency of A. actinomycetemcomitans was 56% between the two sampling methods. A. actinomycetemcomitans was more readily detected by paper point compared with curette collection with a stronger correlation between the two methods in AgP. Subgingival biofilm sampling by curette or paper point does not yield differences in the detection of the three 'red complex' species. However, A. actinomycetemcomitans was more consistently detected by means of paper point collection, which can be crucial in the decision to administer antibiotics as an adjunctive periodontal treatment.

Scope This position paper describes the view adopted by EUCAST on the role of daptomycin in the treatment of serious infections caused by Enterococcus species. Background High-dose daptomycin is considered effective in the treatment of enterococcal bloodstream infection (BSI) and endocarditis, although published clinical experience with the latter condition is limited. Methods EUCAST reviewed the available published data on pharmacokinetics–pharmacodynamics (PK-PD), resistance selection, clinical efficacy and safety for the use of 10–12 mg/kg/day of daptomycin for these conditions, noting that the doses licensed by the European Medicines Agency are only 4–6 mg/kg/day, and only for infections caused by Staphylococcus aureus. Findings and recommendations The PK-PD evidence shows that, even with doses of 10–12 mg/kg/day, it is not possible to treat infections caused by isolates at the upper end of the wild-type distributions of Enterococcus faecalis (with MICs of 4 mg/L) and E. faecium (with MICs of 4 or 8 mg/L). For this reason, and because there are ongoing issues with the reliability of laboratory testing, EUCAST lists daptomycin breakpoints for Enterococcus species as “IE”—insufficient evidence. EUCAST advises increased vigilance in the use of high-dose of daptomycin to treat enterococcal BSI and endocarditis. Additional PK-PD studies and prospective efficacy and safety studies of serious Enterococcal infections treated with high-dose daptomycin may permit the setting of breakpoints in the future.

Adverse effects of pulp and paper mill effluent on fish populations have been well documented in many countries over the last two decades. Some of the initial studies were at mills with conventional chlorine bleaching and no secondary effluent treatment. Following installation of secondary treatment, changes in bleaching technology to elemental chlorine-free bleaching, and other process changes, adverse effects on fish were reduced or eliminated at some mills. Because no two mills are exactly alike, it is difficult to predict adverse impacts of any given mill on fish populations. In 1994, a study of female white sucker (Catostomus commersoni) in the Androscoggin River, Maine, USA, showed induction of mixed function oxidase, reductions in gonad size and plasma estradiol, and an increase in plasma testosterone in fish downstream of discharges from three large bleached kraft pulp and paper mills, and host community municipal sewage treatment plants (STP). After all three mills switched to elemental chlorine-free bleaching in the late 1990s, studies from 2001 to 2003 found that the pattern of reproductive impacts on white sucker populations measured in 1994 was not repeated. In addition, population estimates of white sucker from 2002 to 2003 using mark–recapture techniques found that densities and biomass were well within the range of those of a reference population, and of those reported in the literature for unimpacted populations. Detailed studies immediately above and below each mill/sewage treatment plant showed no evidence of reproductive effects. However, a clear pattern of eutrophication was noted, which increased cumulatively downstream below each mill/STP. Environ. Toxicol. Chem. 2011;30:142–153. © 2010 SETAC

Interferon (IFN)-γ release assays (IGRAs) are probably the most accurate tests for the detection of latent Mycobacterium tuberculosis infection, but IGRAs are labour intensive and the transport of samples over longer distances is difficult. IFN-γ-induced protein (IP)-10 is expressed at 100-fold higher levels than IFN-γ, and IP-10 release assays have comparable performance to IGRAs. The aim of this study was to explore the diagnostic potential of a novel IP-10 release assay based on dried plasma spots (DPS). The presence of IP-10 and IFN-γ was determined in plasma and in DPS by ELISA. Diagnostic algorithms for plasma and DPS tests for IP-10 were developed on a training cohort comprising 60 tuberculosis (TB) patients and 59 healthy controls. Diagnostic accuracy was assessed in a validation cohort comprising 78 TB patients and 98 healthy controls. Plasma was measured in Spain and DPS samples were sent to Denmark using the conventional postal service for analysis. IP-10 was readily detectable in both plasma and DPS, and correlation was excellent (r2 = 0.95). QuantiFERON-TB Gold In-Tube (QFT-TB) and IP-10 in DPS and plasma rendered comparable sensitivity (78%, 82% and 84%, respectively), specificity (100%, 97% and 97%, respectively) and indeterminate rates (p>0.55). The DPS-based IP-10 test has comparable diagnostic accuracy to the QFT-TB and samples can be sent via conventional mail over long distances for analysis without affecting the results.

INTRODUCTION: Human serofrequency of antibodies against Taenia solium antigens was determined and risk factors for cysticercosis transmission were identified. METHODS: Individuals (n=878) from periurban and rural locations of Lages, SC, were interviewed to gather demographic, sanitary and health information. Interviews and blood sample collections by finger prick on Whatman filter paper were performed from August 2004 to May 2005. Observation determined that 850 samples were suitable for analysis and were tested by ELISA using vesicular fluid of Taenia crassiceps heterologous antigen. To ensure the reliability of the results, 77 samples of the dried blood were matched with sera. The reactive samples were submitted to a serum confirmatory immunoblot (IB) test using purified Taenia crassiceps glycoproteins. RESULTS: The ELISA results for the dried blood and serum samples were statistically consistent. ELISA was positive in 186 (21.9%) out of 850 individuals. A group of 213 individuals were asked to collect vein blood for IB (186 with positive result in ELISA and 27 with inappropriate whole blood samples) and 130 attended the request. The IB was positive in 29 (3.4%) out of 850 individuals. A significant correlation (p = 0.0364) was determined among individuals who tested positive in the IB assay who practiced both pig rearing and kitchen gardening. CONCLUSIONS: ELISA with dried blood eluted from filter paper was suitable for cysticercosis population surveys. In Lages, human infection was associated with pig rearing and kitchen gardening. The prevalence index was compatible with other Latin American endemic areas. INTRODUÇÃO: O primeiro levantamento sobre cisticercose humana e identificação dos fatores de risco associados à transmissão, foram realizados em Lages, SC. MÉTODOS: Oitocentos e setenta e sete voluntários de regiões periurbana e rural foram entrevistados e forneceram informações demográficas e condições sanitárias e de saúde. Amostras de sangue foram coletadas por meio de punção digital em papel filtro entre agosto 2004 e maio 2005. Verificou-se que 850 amostras estavam adequadas para análise. No ELISA, utilizou-se o antígeno heterólogo liquido vesicular de Taenia crassiceps. Para assegurar a confiabilidade dos resultados de ELISA, foram pareadas 77 amostras de soro e sangue eluido do papel filtro. A confirmação do diagnóstico sorológico foi feita por immunoblot (IB) com glicoproteínas purificadas de Taenia crassiceps. RESULTADOS: A reatividade de IgG eluída de sangue em papel filtro mostrou-se compatível com a dos soros correspondentes. A triagem por ELISA de 850 indivíduos revelou 186 (21,9%) positivos. De 213 pessoas convidadas a colher soro para IB (186 ELISA positivo e 27 com amostras de sangue total inadequadas), compareceram 130. O IB foi positivo em 29 (3,4%) de 850 amostras. Houve correlação significativa entre IB positivo e a prática de criação de suínos e de horta caseira (p = 0,0364). CONCLUSÕES: ELISA com sangue total em papel filtro mostrou-se adequado para inquéritos populacionais para cisticercose. A transmissão da cisticercose humana na área estudada mostrou correlação com criação suína domestica e horta caseira. A prevalência obtida foi semelhante à relatada em áreas endêmicas da América Latina.

Aedes aegypti (L.) is a major vector of yellow fever, dengue, and Zika viruses, and its management can be difficult, especially in situations where insecticide usage is restricted and resistance is present. Traps and trapping techniques have mostly been used for monitoring populations of adult mosquitoes, but several commercially available traps have been evaluated and used to reduce nuisance populations of adult mosquitoes (Kline 2006). Suppression of Ae. aegypti, in particular, requires a suite of integrated control measures. One measure gaining more attention is to attract and kill gravid females by exploiting their oviposition behavior. Recently, a commercial larval trap for control of Culex mosquito larvae has been developed and marketed. The commercial brand name is My Mosquito Deleter (MMD; Destin, FL). When gravid female Culex mosquitoes lay their eggs in the MMD larval trap filled with water, the larvae after hatching from eggs will fall downward through the MMD's baffle system. The mosquito larvae cannot come to the surface due to the physical barrier from the baffle ring and black cone, resulting in larval mortality. During the preliminary experiment with the original MMD trap full of water, no adult mosquitoes were collected when larval mosquitoes were commonly recorded, because the gravid mosquitoes flew away after they laid their eggs. In order to catch gravid Aedes mosquitoes when they come to the trap to lay their eggs on water within the containers, we modified the MMD trap by placing sticky paper around the inside at the top of the trap (at the water line) after removing the baffle ring and lowering the water level to create an air pocket. The purpose of the study was to investigate the capability of a trap originally designed to trap Culex larvae to attract and kill gravid Ae. aegypti females with a simple and inexpensive modification by adding a piece of black sticky paper and lowering the level of water, in contrast to the unmodified MMD trap with a lower level of water.

The two paper wasps, Polistes dominula and Polistes gallicus, are related species with strongly differing distribution ranges. We investigated thermal tolerance traits (critical thermal limits and metabolic response to temperature) to gain knowledge about physiological adaptations to their local climate conditions and to get evidence for the reasons of P. dominula’s successful dispersion. Body and ambient temperature measurements at the nests revealed behavioural adaptations to microclimate. The species differed clearly in critical thermal minimum (P. dominula −1.4 °C, P. gallicus −0.4 °C), but not significantly in critical thermal maximum of activity (P. dominula 47.1 °C, P. gallicus 47.6 °C). The metabolic response did not reveal clear adaptations to climate conditions. At low and high temperatures, the metabolic rate of P. dominula was higher, and at intermediate temperatures, we determined higher values in P. gallicus. However, the species exhibited remarkably differing thermoregulatory behaviour at the nest. On average, P. gallicus tolerated a thoracic temperature up to ~41 °C, whereas P. dominula already tried at ~37 °C to keep the thorax below ambient temperature. We suggest this to be an adaptation to the higher mean ambient temperature we measured at the nest during a breeding season. Although we determined for P. dominula a 0.5 °C larger thermal tolerance range, we do not presume this parameter to be solely responsible for the successful distribution of P. dominula. Additional factors, such as the thermal tolerance of the queens could limit the overwintering success of P. gallicus in a harsher climate.

The data pertaining to the COVID‐19 pandemic has been rapidly evolving since the first confirmed case in December 2019. This review article presents a comprehensive analysis of the current data in relation to COVID‐19 and its effect on pregnant women, including symptoms, disease severity and the risk of vertical transmission. We also review the recommended management of pregnant women with suspected or confirmed COVID‐19 and the various pharmacological agents that are being investigated and may have a role in the treatment of this disease. At present, it does not appear that pregnant women are at increased risk of severe infection than the general population, although there are vulnerable groups within both the pregnant and nonpregnant populations, and clinicians should be cognizant of these high‐risk groups and manage them accordingly. Approximately 85% of women will experience mild disease, 10% more severe disease and 5% critical disease. The most common reported symptoms are fever, cough, shortness of breath and diarrhea. Neither vaginal delivery nor cesarean section confers additional risks, and there is minimal risk of vertical transmission to the neonate from either mode of delivery. We acknowledge that the true effect of the virus on both maternal and fetal morbidity and mortality will only be evident over time. We also discuss the impact of social isolation can have on the mental health and well‐being of both patients and colleagues, and as clinicians, we must be mindful of this and offer support as necessary.

The acute and chronic effects of secondary-treated effluent from a New Zealand pulp and paper mill were assessed using both long-term adult and early life stage (ELS) laboratory exposures of rainbow trout. The relative impact of maternal exposure versus ELS exposure was assessed by a comparison of directly exposed eggs and larvae with the eggs and larvae of exposed adult trout that were reared in reference water. Rainbow trout were exposed to a secondary-treated mixed thermomechanical/bleached kraft mill effluent at a concentration of 15% or to reference water from the egg through to 320-d-old juveniles. The 2 adult rainbow trout exposures were undertaken with nominal concentrations of 10% and 12% treated effluent, respectively. There was no marked effect of water hardening with 15% effluent on fertility or survival of eggs to 16 d. In a subsequent exposure (with hardening in reference water), no significant effects were found on mortality to hatch, time to hatch, length at hatch, mortality to swim-up, mortality to 320 d, or deformity rate at hatch. At experimental termination (320 d), direct-exposed juveniles had smaller livers and reduced condition factor, likely due to differences in food consumption. In 2 subsequent consecutive experiments, exposure of adult trout to 10% and 12% effluent for 2 mo, followed by incubation of the fertilized eggs in reference water, produced no impact on fertility, survival to hatch, survival to swim-up, or length and weight of fry at swim-up. Exposure of adult trout to 12% treated effluent for 8 mo prior to egg fertilization also did not result in differing rates of fertility, mortality to hatch or mortality to swim-up. However, the 8-mo maternal exposure did result in swim-up fry that were significantly shorter and weighed less than the reference swim-up fry. This difference was directly attributable to smaller eggs in the 8-mo-exposed female trout. These results demonstrate that this pulp and paper mill effluent is more likely to elicit indirect impacts on progeny size through chronic exposure of adults to effluent during gonadal recrudescence rather than through direct exposure of early life stages to effluent.

The attack phase of the visceral leishmaniasis (VL) elimination program in Bangladesh aims to decrease the burden of VL incidence from close to 20 cases to less than one case per 10,000 at sub-district level. The consolidation phase will aim to confirm no increase in VL in endemic areas through active surveillance. During this phase, a reliable diagnostic tool for mass screening is required. Here, we report the diagnostic sensitivity and specificity of a filter paper-based agglutination test (FP-DAT) for diagnosis of VL in patients admitted to an upazila health complex in Mymensingh, a VL-endemic region of Bangladesh. The sensitivity of both the conventional direct agglutination test (DAT) and FP-DAT were 100% and 96%, respectively. The specificity of both assays was 100%. However, when the performances of the two assays were compared using McNamar's test, neither the sensitivity nor the specificity of the FP-DAT differed significantly from conventional DAT.

The collection of subgingival plaque samples with paper points is time-consuming and accident-sensitive. However, the collection of saliva is simple and contains pathogens of all intraoral surfaces. The aim of this study is to investigate whether a sampling strategy with mouthrinse (mouthrinse sample [MSP]; test) leads to results comparable with standard sampling method (multiple site test from the deepest pocket of each quadrant [MT4]; control).In 50 patients with periodontitis, subgingival plaque was sampled from the deepest pocket of each quadrant by using paper points and by gaining saliva with saline mouthrinse. Analysis was performed using a commercially available polymerase chain reaction test for 11 periodontal pathogens.Detection frequency of Aggregatibacter actinomycetemcomitans (MT4/MSP: 42%/36%), Porphyromonas gingivalis (78%/66%), Tannerella forsythia (98%/84%), Treponema denticola (94%/74%), Parvimonas micra (86%/62%), Campylobacter rectus (90%/76%), Eubacterium nodatum (64%/30%), Prevotella intermedia (58%/54%), and Eikenella corrodens (90%/82%) was higher with MT4 than MSP. For Fusobacterium nucleatum (100%/100%), there was no difference between test and control. Only detection frequency of Capnocytophaga species (68%/74%) was higher with MSP than MT4. Differences were significant for P. gingivalis, T. forsythia, T. denticola, P. micra, C. rectus, and E. nodatum.There is no significant difference between MT4 and MSP for detection frequency of key pathogen A. actinomycetemcomitans. Key pathogens P. gingivalis, T. forsythia, T. denticola, P. micra, C. rectus, and E. nodatum show statistically higher detection frequencies with MT4.

In cooperatively breeding vertebrates, the existence of individuals that help to raise the offspring of non-relatives is well established, but unrelated helpers are less well known in the social insects. Eusocial insect groups overwhelmingly consist of close relatives, so populations where unrelated helpers are common are intriguing. Here, we focus on Polistes dominula— the best-studied primitively eusocial wasp, and a species in which nesting with non-relatives is not only present but frequent. We address two major questions: why individuals should choose to nest with non-relatives, and why such individuals participate in the costly rearing of unrelated offspring. Polistes dominula foundresses produce more offspring of their own as subordinates than when they nest independently, providing a potential explanation for co-founding by non-relatives. There is some evidence that unrelated subordinates tailor their behaviour towards direct fitness, while the role of recognition errors in generating unrelated co-foundresses is less clear. Remarkably, the remote but potentially highly rewarding chance of inheriting the dominant position appears to strongly influence behaviour, suggesting that primitively eusocial insects may have much more in common with their social vertebrate counterparts than has commonly been thought.

Adult workers of some social insect species show dramatic behavioral changes as they pass through a sequence of task specializations. In the paper wasp, Polybia aequatorialis, female workers begin adult life within the nest tending brood, progress to maintaining and defending the nest exterior, and ultimately leave the nest to forage. The mushroom body (MB) calyx neuropil increases in volume as workers progress from in-nest to foraging tasks. In other social Hymenoptera (bees and ants), MB Kenyon cell dendrites, axons and synapses change with the transition to foraging, but these neuronal effects had not been studied in wasps. Furthermore, the on-nest worker of Polybia wasps, an intermediate task specialization not identified in bees or ants, provides the opportunity to study pre-foraging worker class transitions. We asked whether Kenyon cell dendritic arborization varies with the task specialization of Polybia workers observed in the field near Monteverde, Costa Rica. Golgi-impregnated arbors in the lip and collar calyces, which receive a predominance of olfactory and visual input, respectively, were quantified using Sholl's concentric circles and a novel application of virtual spherical probes. Arbors of the lip varied in a manner reminiscent of honeybees, with foragers having the largest and in-nest workers having the smallest arbors. In contrast, arbors of the collar were largest in foragers but smallest in on-nest workers. Thus, progression through task specializations in P. aequatorialis involves subregion specific dendritic growth and regression in the MB neuropil. These results may reflect the sensitivity of Kenyon cell dendritic structure to specialization dependent social and sensory experience.

Explaining the evolution of helping behaviour in the eusocial insects where non-reproductive ('worker') individuals help raise the offspring of other individuals ('queens'), remains one of the most perplexing phenomena in the natural world. Polistes paper wasps are popular study models, as workers retain the ability to reproduce: such totipotency is likely representative of the early stages of social evolution. Polistes is thought to have originated in the tropics, where seasonal constraints on reproductive options are weak and social groups are effectively perennial. Yet, most Polistes research has focused on non-tropical species, where seasonality causes family groups to disperse; cofoundresses forming new colonies the following spring are often unrelated, leading to the suggestion that direct fitness through nest inheritance is key in the evolution of helping behaviour. Here we present the first comprehensive genetic study of social structure across the perennial nesting cycle of a tropical Polistes - Polistes canadensis. Using both microsatellites and newly-developed single-nucleotide polymorphism (SNP) markers we show that adult cofoundresses are highly related, and that brood production is monopolised by a single female across the nesting cycle. Non-reproductive cofoundresses in tropical Polistes therefore have the potential to gain high indirect fitness benefits as helpers from the outset of group formation, and these benefits persist through the nesting cycle. Direct fitness may have been less important in the origin of Polistes sociality than previously suggested. These findings stress the importance of studying a range of species with diverse life-history and ecologies when considering the evolution of reproductive strategies.

Regulatory pathways in solitary species provide the raw materials for the evolution of sociality. Therefore, comparing the mechanisms that mediate reproductive plasticity in social species and their solitary ancestors can provide insight into the evolutionary origin of sociality. In many solitary insects, the effect of juvenile hormone (JH) on fertility is mediated through the fat body; individuals in good physical condition show a stronger fertility response to JH than individuals in poor physical condition. Here, we test whether a similar, condition-dependent JH response mediates fertility in workers of the primitively eusocial Polistes dominulus wasps. We test how body weight, JH, and adult nutrition influence worker ovarian development. Both JH-treatment and adult nutrition dramatically increased ovarian development. Body weight also influenced ovarian development, as large workers developed more eggs than smaller workers. Body weight and fat are strongly linked in P. dominulus workers, so these results suggest that the fat-dependent JH responsiveness common in solitary insects is conserved in social wasps. The simple, ancestral relationship between reproductive investment and physical condition may facilitate cooperation by allowing workers to adaptively allocate energy into reproduction based on their probability of successfully becoming a queen.

SUMMARY Foundresses and workers of Polistes paper wasps show slight morphological and physiological differences. However, after the emergence of the workers, the castes can be readily discriminated by their behaviour: the dominant foundress is the principal egg-layer, whereas workers perform different tasks linked to colony development. Previous studies have demonstrated in this genus that defence of the colony by the workers is more effectively carried out by a collective response elicited by venom volatiles used as alarm pheromones. In the present study, gas chromatography–mass spectrometry analyses of the venom volatiles of foundresses and workers of Polistes dominulus (Christ) show predominantly quantitative differences. Spiroacetals, mainly(E,E)-2,8-dimethyl-1,7-dioxaspiro[5.5]undecane, are significantly higher in the venom volatiles fraction of workers, whereas the amount of N-(3-methylbutyl)acetamide is almost double in foundresses. On the basis of the chemical results, behavioural assays were performed on fifteen field colonies to test the alarm response of the resident wasps to venom extracts from foundresses and workers. Our behavioural results suggest that worker venom has a stronger alarm effect on the colonies than that of the foundresses, which seems unable to elicit the complete alarm response ending with a final attack and sting. The venom volatiles of P. dominulusworkers serve mainly to alarm the colony whilst those of foundresses may also be linked to additional functions related to conspecific interactions.

To establish a dominance order, social animals often rely on indicators of fighting to avoid costly aggressive encounters. In some species, individuals use colour patterns to signal their social status. Recent studies claimed that facial markings in the eusocial paper waspPolistes dominulusare status badges that allow co-foundresses to form a linear hierarchy based on individual quality. Here, we evaluated facial patterns in natural populations ofP. dominulus, in its native range, to observe whether the marks reflect overall wasp quality in different contexts. We used the same measures of clypeus patterns used by earlier studies, but did not find that they functioned as status badges. Our analyses showed no evidence that visual markers are related to: (i) size, (ii) probability of surviving winter, (iii) social rank in spring associations, or (iv) health status (assessed by the presence of strepsipteran endoparasites). Size, however, is important. Larger wasps are more likely to survive the winter and to acquire the dominant position in spring associations. Larvae infected with endoparasites become smaller adult wasps. These findings suggest that body size is a reliable quality indicator on which wasps build their social networks, and that clypeus patterning is not involved.

Several studies have shown that differences in the relative abundance of cuticular hydrocarbons occur between reproductives and non-reproductives in many social insects. These differences also exist between Polistes dominulus foundresses and their first emerged daughters (usually indicated as workers), but they gradually disappear when workers from orphaned colonies develop their ovaries and assume a reproductive role in the colony. However, hydrocarbons are not the exclusive components of cuticular layer of wasps. Mass-spectrometry analysis of cuticular methanol extracts from Polistes paper wasps showed a complex pattern of polar substances, partly or totally proteinaceous in nature. We found that these compounds, ranging from 918 to 2679 Da, showed a clear caste differentiation between foundresses and their first emerged daughters (usually indicated as "workers"), both in queen-right and in orphaned colonies. Conversely to hydrocarbons, workers from orphaned colonies maintain a significant difference from foundresses in the pattern of the medium molecular weight (MW) polar compounds obtained by MALDI-TOF. On the basis of such results we hypothesize that a reliable cue to identify foundresses from daughters, and not only their reproductive status, may exist in Polistes wasps. Although the great majority of previous work on social insect communication has focused on cuticular hydrocarbons, our findings suggest that the medium MW component of cuticular substances may be involved in recognition.

Female mosquitofish exposed to pulp and paper mill effluent (PME) in the Fenholloway River, Florida, USA have masculinized secondary sex characteristics and altered aromatase enzyme activity. We and others have shown that the Fenholloway River PME contains androgenic and progestogenic substance(s). The present study was designed to test the hypothesis that the development and reproductive health of PME-exposed Fenholloway River mosquitofish are altered compared to mosquitofish living in Econfina River, which is the reference site. Fish were collected on a single day from both sites in June and August 1999 and January and June 2000. We compared standard length, anal fin length and segment number; body, liver, and gonad mass; and number of eggs and embryos from Fenholloway and Econfina River mosquitofish. The data were analyzed collectively for generalized site effect, for site effects during reproductive and nonreproductive seasons, and for repeatability of site effects between years. Mosquitofish exposed to PME in the Fenholloway River were generally smaller in length and mass, anal fin segment number was greater, and the number of embryos, but not oocytes, was significantly decreased compared to the reference site fish. Anal fin length and segment number and liver and testis masses were generally greater in Fenholloway compared to the Econfina River males. The importance of this study is that we have documented masculinized development and decreased embryo production in PME-exposed mosquitofish and that these site effects are generally consistent across seasons and between years.

Several environmental contaminants have been found in paper mill effluent including substances with androgenic properties. The presence of androgenic substances in paper mill effluent was also indicated in the present study which demonstrated masculinized anal fins of female mosquitofish from the paper mill effluent contaminated Fenholloway River. In addition, when compared to the nearby Econfina River, which does not receive paper mill effluent, fewer females from the contaminated river were pregnant, they were smaller and their estradiol concentration exhibited greater variation. Males from the Fenholloway River and the Econfina River had similar sperm counts, but the testes were larger and greater variation in testosterone concentration was observed in male fish from the Fenholloway River. In males and females from the Fenholloway River, liver weights were increased and computer-aided behavior analysis demonstrated a reduction in their social behavior when compared to reference fish. We conclude that a number of sexual characteristics were affected in mosquitofish living in the paper mill contaminated Fenholloway River, with possible adverse effects on the reproduction of this population.

Although gender mainstreaming has been long recognized as a strategy for addressing gender inequalities and associated negative health outcomes; its implementation has remained a challenge, even in the area of prevention of mother to child transmission of HIV (PMTCT). Despite recognition of gender in Tanzania’s political arena and prioritization of PMTCT by the health sector, there is very little information on how well gender has been mainstreamed into National PMTCT guidelines and organizational practices at service delivery level. Using a case study methodology, we combined document review with key informant interviews to assess gender mainstreaming in PMTCT on paper and in practice in Tanzania. We reviewed PMTCT policy/strategy documents using the WHO’s Gender Responsive Assessment Scale (GRAS). The scale differentiates between level 1 (gender unequal), 2 (gender blind), 3 (gender sensitive), 4 (gender specific), and 5 (gender transformative). Key informant interviews were also conducted with 26 leaders purposively sampled from three government health facilities in Mwanza city to understand their practices. The gender responsiveness of PMTCT policy/strategy documents varies, with some being at GRAS level 3 (gender sensitive) and others at GRAS level 4 (gender specific). Those which are gender sensitive indicate gender awareness, but no remedial action is developed; while those which are gender specific go beyond indicating how gender may hinder PMTCT to highlighting remedial measures, such as the promotion of couple counselling and testing for HIV. In addition, interviews on organizational processes and practices suggested that there has been little attention to the holistic integration of gender in the delivery of PMTCT services. The study has revealed limited integration of gender concerns in PMTCT policy documents. Similarly, health facility leader responses indicate perspectives and practices that pay little attention to the holistic integration of gender in the delivery PMTCT services.

Social wasps from temperate zones have clear annual colony cycles, and the young queens hibernate during winter. In the subtropics, the only previously reported evidence for the existence of "hibernation" is the facultative winter aggregations of females during harsh climate conditions. As in temperate-zone species analyzed so far, we show in this study that in the paper wasp, Polistes versicolor, a subtropical species, body size increases as an unfavorable season approaches. Our morphological studies indicate that larger females come from winter aggregations--that is, they are new queens. Multivariate analyses indicate that size is the only variable analyzed that shows a relationship to the differences. Given the absence of a harsh climate, we suggest that the occurrence of winter aggregations in tropical P. versicolor functions to allow some females to wait for better environmental conditions to start a new nest, rather than all being obliged to start a new nest as soon as they emerge.

Summary The prevalence of lymphatic filariasis was estimated by PCR-based pool screening of night blood collected from 865 individuals living in ten areas endemic for Wuchereria bancrofti , Brugia malayi or B. timori in Indonesia. A total of 232 microfilaraemics were identified by filtration of 1 ml of blood. The microfilaria (mf) prevalence ranged from 6% to 54%, and the mf density in microfilaraemics ranged from 1 mf/ml to 6028 mf/ml. PCR assays both for W. bancrofti or Brugia spp. detected a single mf present on a 30 μl dried filter paper blood spot. One hundred and seventy-eight pools of five blood spots in each pool (pool-5) were tested by PCR and 101 (57%) pools were positive. When pool size was increased to 10 spots per pool (pool-10), 65 (70%) of 93 pools were positive. Pearson's correlation and linear regression showed a strong correlation between filtration and pool screen PCR results for pool-10 ( r = 0.835) and pool-5 ( r = 0.695). Based on the determination coefficient ( R ), the results of pool-10 PCR ( R = 0.697) gave a better prediction compared with pool-5 PCR ( R = 0.483). This study suggests that pool screen PCR may be a useful tool for monitoring the Global Program to Eliminate Lymphatic Filariasis.

Studies have shown that female mosquitofish living downstream of a paper mill located on the Fenholloway River, Florida, have masculinized secondary sex characteristics, including altered anal fin development and reproductive behavior. Masculinization can be caused by exposure to androgens in the water or from an alteration in aromatase activity in the fish. We hypothesized that aromatase activity would be inhibited by a component(s) of the paper mill effluent. Aromatase inhibition could masculinize the hormonal profile and, subsequently, secondary sex characteristics of the exposed females. Therefore, we predicted that ovarian and brain aromatase activity would be lower in the female mosquitofish from the Fenholloway River compared with the reference site, the Econfina River. Adult females were collected and standard length, body mass, anal fin length, and segment number were measured. Ovarian and brain aromatase activity were determined using a tritiated water assay. Fenholloway females had masculinized anal fin development as indicated by an increase in the number of segments in the longest anal fin ray (p < 0.0001), yet the length of the ray did not differ between sites (p = 0.95). Fenholloway females exhibited higher ovarian (p = 0.0039) and brain (p = 0.0003) aromatase activity compared with reference site fish. These data do not support aromatase inhibition as the mechanism for masculinization, suggesting that the masculinization of the Fenholloway female mosquitofish is due to androgenic contaminants. Future studies should examine the relationship between aromatase enzyme activity and exposure to environmental androgens.

The objective of this study was to evaluate the effects of bleached/unbleached kraft mill effluents (B/UKME) on the reproductive parameters of free-ranging Florida largemouth bass (Micropterus salmoides floridanus). The reproductive parameters measured included gonadosomatic index (GSI), histological evaluation of gonads, and plasma concentrations of vitellogenin (VTG), 17beta-estradiol, and 11-ketotestosterone (11-KT). Hepatic ethoxyresorufin-O-deethylase (EROD) activity was measured as a marker of exposure to cytochrome P450-inducing agents in these effluents. Endpoints were compared among adult bass sampled from tributary and mainstream effluent-contaminated and reference sites. Females sampled from the site closest to the mill outfall had a significant five-fold increase in EROD activity compared to bass sampled from reference streams. Although sex hormones were significantly reduced in bass from exposed sites, there were no differences in VTG and GSI across sites. The absence of organism-level responses was probably not related to a lack of sensitivity, as previous studies in our laboratory have shown that bass exposed to these effluents exhibit changes in GSI and in other measures associated with reproductive success. In females, inverse relationships were observed between VTG and GSI and EROD activity. These relationship, however, were not consistent within all of the sites studied. Collectively, our findings indicate that hepatic EROD induction is an effective marker of B/UKME exposure in largemouth bass and that it might be associated with antiestrogenic effects in this species.

Compared to our extensive knowledge about the navigation and homing abilities of ants and bees, we know rather little about these phenomena in social wasps. Here, we report the homing abilities of the tropical primitively eusocial wasp Ropalidia marginata and the factors that affect their homing success. To determine from how far these wasps can return to their nests, we transported foragers blindfold and released them at gradually increasing distances from their nests in four cardinal directions. Their homing success was determined by checking their presence on their nests on three consecutive nights. All foragers (56 individuals, 115 releases) returned back from an area of 0.73 +/- A 0.25 km(2) on the day of release (minimal homing area), whereas 83.8 % of the foragers (217 individuals, 420 releases) returned when we enlarged the area of release to 6.22 +/- A 0.66 km(2) around their nests (maximal homing area). Of 66 releases, no wasps returned from beyond the maximal homing area. The minimal homing area might be familiar to the foragers because they probably routinely forage in this area and the maximal homing area represents the maximum distances from which the wasps are capable of returning to their nests, with or without familiarity.

The distinction between worker and reproductive castes of social insects is receiving increased attention from a developmental rather than adaptive perspective. In the wasp genus Polistes, colonies are founded by one or more females, and the female offspring that emerge in that colony are either non-reproducing workers or future reproductives of the following generation (gynes). A growing number of studies now indicate that workers emerge with activated reproductive physiology, whereas the future reproductive gynes do not. Low nourishment levels for larvae during the worker-rearing phase of the colony cycle and higher nourishment levels for larvae when gynes are reared are now strongly suspected of playing a major role in this difference. Here, we present the results of a laboratory rearing experiment in which Polistes metricus single foundresses were held in environmental conditions with a higher level of control than in any previously published study, and the amount of protein nourishment made available to feed larvae was the only input variable. Three experimental feeding treatments were tested: restricted, unrestricted, and hand-supplemented. Analysis of multiple response variables shows that wasps reared on restricted protein nourishment, which would be the case for wasps reared in field conditions that subsequently become workers, tend toward trait values that characterize active reproductive physiology. Wasps reared on unrestricted and hand-supplemented protein, which replicates higher feeding levels for larvae in field conditions that subsequently become gynes, tend toward trait values that characterize inactive reproductive physiology. Although the experiment was not designed to test for worker behavior per se, our results further implicate activated reproductive physiology as a developmental response to low larval nourishment as a fundamental aspect of worker behavior in Polistes.

To learn the evolutionary trajectories of caste differentiation in eusocial species is a major goal of sociobiology. We present an explanatory framework for caste evolution in the eusocial wasp genus Polistes (Vespidae), which is a model system for insect eusocial evolution. We hypothesize that Polistes worker and gyne castes stem from two developmental pathways that characterized the bivoltine life cycle of a solitary ancestor. Through individual-based simulations, we show that our mechanistic framework can reproduce colony-level characteristics of Polistes and, thereby, that social castes can emerge from solitary regulatory pathways. Our explanatory framework illustrates, by specific example, a changed perspective for understanding insect social evolution.

Background: In vitro stimulation of whole blood or isolated blood cells with specific antigens is used for several purposes. Immediately following incubation with antigens, samples have to be centrifuged to stop the reactions by remaining cells and the supernatant refrigerated or analysed directly to preserve the analytes of interest, which makes samples difficult to prepare outside laboratories. We have tested whether spotting whole blood on filter paper after activation can be used in one of the tests for Mycobacterium tuberculosis infection (MTI), the QuantiFERON®-TB Gold In Tube test (QFT), where the spotting technique can make it suitable for use in locations without facilities like a centrifuge and a refrigerator. Materials and methods: Samples from 22 individuals undergoing screening for MTI and 10 healthy controls were incubated, centrifuged and IFN-γ measured by Enzyme-linked immunosorbent assay (ELISA), as described in the kit insert. In parallel, activated blood was spotted on filter paper (Schleicher & Schuell) and dried. The dried blood spot samples were analysed for 21 inflammatory markers with an in-house assay based on Luminex technology. Results: Our multiplex measurements of inflammatory markers in samples from suspected MTI patients confirmed the IFN-γ findings in the QFT. IL-2, GM-CSF, IL-5, and IL-1β were also found as useful markers for MTI. We were not able to distinguish between active tuberculosis and latent MTI. Conclusion: Applying blood on filter paper after incubation makes in vitro stimulation tests feasible in locations where heat and electricity is unavailable.

Sexually selected signals are common in many animals, though little reported in social insects. We investigated the occurrence of male visual signals mediating the dominance relationships among males and female choice of sexual partner in the paper wasp Polistes simillimus. Males have three conspicuous, variable and sexually dimorphic traits: black pigmentation on the head, a pair of yellow abdominal spots and body size differences. By conducting behavioral assays, we found that none of the three visual traits are associated with male-male dominance relationship. However, males with higher proportion of black facial pigmentation and bigger yellow abdominal spots are more likely chosen as sexual partners. Also, after experimentally manipulating the proportion of black pigment on males' face, we found that females may evaluate male facial coloration during the choice of a sexual partner. Thus, the black pigmentation on P. simillimus male's head appears to play a role as a sexually selected visual signal. We suggest that sexual selection is a common force in Polistes and we highlight the importance of this group as a model for the study of visual communication in insects.

The present study aimed at comparing the trypanosome specific 18S-PCR-RFLP using samples stored either on Whatman filter papers (PCR-RFLP-fp) or in a commercial cell lysis and DNA protecting buffer (PCR-RFLP-pb) with the haematocrit centrifugation technique (HCT), a method widely used for the diagnosis of African Animal Trypanosomosis. Out of 411 head of cattle, 49 (11.92%) (CI=8.95-15.45) scored positive for the presence of trypanosomes by HCT whereas 75 (18.25%) (CI=14.63-22.33) and 124 (30.17%) (CI=25.77-34.86) scored positive using PCR-RFLP-fp and PCR-RFLP-pb, respectively. Out of the 49 positives by HCT, 14 (28.57%) (CI=16.58-43.26) and 28 (57.14%) (CI=42.21-71.18) were concordant by PCR-RFLP-fp and PCR-RFLP-pb, respectively. None of the PCR techniques detected parasites from the Trypanozoon group. Although HCT detected more cases of Trypanosoma vivax (33), species identification using PCR-RFLP-fp and PCR-RFLP-pb were significantly different (p0.001) from the HCT technique. The use of DNA protective buffer is thus recommended as the output of the PCR-RFLP-pb is improved and the risk of contamination between samples is reduced.

Different ways of eluting IgG antibodies against Toxoplasma gondii from blood spots on PKU filter papers from newborns were examined, and the eluate was analysed by Enzyme Immunosorbent Assay (EIA). 10-mm diameter discs (method A), and 3-mm discs (method B) were eluted overnight, and 3-mm discs were eluted directly for 2 hours in the EIA wells (method C). Two groups of paired samples were tested: blood spots on the PKU card from the newborn, cord blood, and a venous blood sample from the mother collected shortly after delivery (Group 1, 191 samples); or PKU cards from the newborns and serum collected from the mother during the first trimester (Group 2, 426 samples). The best results were found for Group 1; method A, a sensitivity of 100% (63/63), and a specificity of 100% (128/128) (r = 0.934, P < 0.0001), and for Group 2; method C, a sensitivity of 100% (125/125), and a specificity of 100% (301/301) (r = 0.822, P < 0.0001). When method C, was used on 188 samples stored for 16-18 month at -20 degrees C, a sensitivity of 100% (62/62) and a specificity of 99.2% (125/126) (r = 0.913, P < 0.0001) were found. The PKU filter paper method is reliable and reproducible, and can be used in screening projects based on analysis of neonatal samples, and in epidemiological studies on toxoplasmosis in pregnant women.

Two experiments using rainbow trout (Oncorhynchus mykiss) were conducted to examine the combined effects of energy intake as manipulated by ration and pulp and paper mill effluent exposure over either one, or two consecutive reproductive cycles. This study demonstrated that the level of energy intake affected the full range of measured parameters from energy allocation to somatic growth and the gonadal development, steroid production and hematology. Increasing ration level expectedly increased growth, condition, liver and gonad size. Female trout in the higher ration treatments produced more follicles and had larger eggs, investing the same relative proportion of total energy into ovarian development. Sex steroid levels and hematological parameters were also positively influenced by increasing ration level in males and females. By far, the most dramatic impact of reduced ration on reproduction was to substantially reduce the frequency of sexually maturing fish. The effects of effluent exposure were not as marked as those linked to ration level and typically did not manifest unless fish were exposed through two consecutive reproductive cycles. The physiological effects of pulp and paper effluent exposure observed in these experiments were not consistent between the two experiments conducted herein, nor were they consistent with previously observed impacts in similar experiments with this effluent. Effluent exposure over one reproductive cycle did not impact physiological parameters in trout. However, when effluent exposure was maintained over two reproductive cycles, a new pattern of effluent response emerged including increased condition factor in both sexes, a decrease in the potential ability of the blood of females to transport oxygen, and increased sex steroids and reproductive investment in males. Effluent was also observed to cause reduced growth in male trout over two years. The effects of ration on gonad and liver size were far more obvious and consistent when a longer exposure was employed, thus, it appears to take more than one full year for energy intake changes to be reflected in those particular physiological endpoints.