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Insects have a rich diversity of RNA viruses that can either cause acute infections or persist in host populations without visible symptoms. The melon fly, Zeugodacus cucurbitae (Tephritidae) causes substantial economic losses through infestation of diverse cucurbit and other crops. Of Indomalayan origin, it is now established in many tropical regions of the world. The virome diversity of Z. cucurbitae is largely unknown across large parts of its distribution, including the Indian subcontinent. We have analysed three transcriptomes each of one field-collected and one laboratory-reared Z. cucurbitae population from Bangalore (India) and discovered genomes of ten putative RNA viruses: two sigmaviruses, one chimbavirus, one cripavirus, one noda-like virus, one nora virus, one orbivirus, one partiti-like virus, one sobemovirus and one toti-like virus. Analysis of the only available host genome of a Hawaiian Z. cucurbitae population did not detect host genome integration of the detected viruses. While all ten viruses were found in the Bangalore field population only seven were detected in the laboratory population, indicating that these seven may cause persistent covert infections. Using virus-specific RNA-dependent RNA polymerase gene primers, we detected nine of the RNA viruses with an overall low variant diversity in some but not all individual flies from four out of five Indian regions. We then screened 39 transcriptomes of Z. cucurbitae laboratory populations from eastern Asia (Guangdong, Hainan, Taiwan) and the Pacific region (Hawaii), and detected seven of the ten virus genomes. We found additional genomes of a picorna-like virus and a negev-like virus. Hawaii as the only tested population from the fly's invasive range only had one virus. Our study provides evidence of new and high RNA virus diversity in Indian populations within the original range of Z. cucurbitae, as well as the presence of persistent covert infections in laboratory populations. It builds the basis for future research of tephritid-associated RNA viruses, including their host effects, epidemiology and application potential in biological control., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Objective: To describe the outcomes (change in functional independence and discharge disposition) of patients who after liver transplantation received acute inpatient rehabilitation in a freestanding rehabilitation hospital., Design: A retrospective chart review was conducted of patients admitted to an acute inpatient rehabilitation hospital within 6 months of undergoing liver transplantation between January 2014 and December 2018. Change in function from rehabilitation admission to discharge was measured using FIM Change and FIM Efficiency., Setting: A freestanding rehabilitation hospital., Participants: 107 patients who underwent acute inpatient rehabilitation at a freestanding rehabilitation hospital within 6 months after liver transplantation who met inclusion criteria (N=107). Most were men (71.96%), and the mean age of the patient population was 62.15 years., Interventions: Acute inpatient rehabilitation consisting of at least 3 hours of therapy 5 days a week split between physical therapy, occupational therapy, and speech language pathology services., Main Outcome Measure: FIM Change, FIM Efficiency, Discharge Disposition., Results: Participants were found to have statistically significant positive FIM Change ( P <.00001) and FIM Efficiency ( P <.00001). The mean FIM Change and Efficiency were 35.7±11.8 and 2.4±1.0, respectively. 83.2% (n = 89) were ultimately discharged to the community., Conclusion: Acute inpatient rehabilitation provides patients who have received a liver transplant with the opportunity to measurably improve their function and independence, with most patients being able to return home., (© 2024 The Authors.)

Insect-specific viruses (ISVs) can affect insect health and fitness, but can also interact with other insect-associated microorganisms. Despite this, ISVs are often studied in isolation from each other, in laboratory populations. Consequently, their diversity, prevalence and associations with other viruses in field populations are less known, yet these parameters are important to understanding virus epidemiology. To help address this knowledge gap, we assessed the diversity, prevalence and coinfections of three ISVs (horizontally transmitted cripavirus, biparentally transmitted sigmavirus and maternally transmitted iflavirus) in 29 field populations of Queensland fruit fly, Australia's most significant horticultural pest, in the context of their different transmission modes. We detected new virus variant diversity. In contrast to the very high virus prevalence in laboratory populations, 46.8% of 293 field flies carried one virus and 4.8% had two viruses. Cripavirus and sigmavirus occurred in all regions, while iflavirus was restricted to subtropical and tropical regions. Cripavirus was most prevalent (37.5%), followed by sigmavirus (13.7%) and iflavirus (4.4%). Cripavirus coinfected some flies with either one of the two vertically transmitted viruses. However, sigmavirus did not coinfect individuals with iflavirus. Three different modelling approaches detected negative association patterns between sigmavirus and iflavirus, consistent with the absence of such coinfections in laboratory populations. This may be linked with their maternal transmission and the ineffective paternal transmission of sigmavirus. Furthermore, we found that, unlike sigmavirus and iflavirus, cripavirus load was higher in laboratory than field flies. Laboratory and mass-rearing conditions may increase ISV prevalence and load due to increased transmission opportunities. We conclude that a combination of field and laboratory studies is needed to uncover ISV interactions and further our understanding of ISV epidemiology., (© 2023 The Authors. Molecular Ecology published by John Wiley & Sons Ltd.)

Microbiomes play vital roles in insect fitness and health and can be influenced by interactions between insects and their parasites. Many studies investigate the microbiome of free-living insects, whereas microbiomes of endoparasitoids and their interactions with parasitised insects are less explored. Due to their development in the constrained environment within a host, endoparasitoids are expected to have less diverse yet distinct microbiomes. We used high-throughput 16S rRNA gene amplicon sequencing to characterise the bacterial communities of Dipterophagus daci (Strepsiptera) and seven of its tephritid fruit fly host species. Bacterial communities of D. daci were less diverse and contained fewer taxa relative to the bacterial communities of the tephritid hosts. The strepsipteran's microbiome was dominated by Pseudomonadota (formerly Proteobacteria) (> 96%), mainly attributed to the presence of Wolbachia, with few other bacterial community members, indicative of an overall less diverse microbiome in D. daci. In contrast, a dominance of Wolbachia was not found in flies parasitised by early stages of D. daci nor unparasitised flies. Yet, early stages of D. daci parasitisation resulted in structural changes in the bacterial communities of parasitised flies. Furthermore, parasitisation with early stages of D. daci with Wolbachia was associated with a change in the relative abundance of some bacterial taxa relative to parasitisation with early stages of D. daci lacking Wolbachia. Our study is a first comprehensive characterisation of bacterial communities in a Strepsiptera species together with the more diverse bacterial communities of its hosts and reveals effects of concealed stages of parasitisation on host bacterial communities., (© 2023. The Author(s).)

Infections of insects with insect-specific RNA viruses are common and can affect host fitness and health. Previously, persistent RNA virus infections were detected in tephritid fruit flies, including the Queensland fruit fly (Bactrocera tryoni), Australia's most significant horticultural pest. Their transmission modes and efficiency are unclear yet may influence virus epidemiology in field and laboratory populations. Using standard RT-PCR and RT-qPCR we detected iflavirus, cripavirus and sigmavirus in five laboratory populations recently established with field-collected B.tryoni. Virus absence in some individuals suggested that virus transmission is incomplete. Random virus segregation in an isofemale experiment resulted in the establishment of isofemale lines with and without iflavirus and cripavirus. In infected lines, viral loads normalised against host gene transcripts were variable, but did not differ between pupae and adults. Iflavirus and cripavirus were transmitted horizontally, with viruses detected (including at low viral loads) in many previously uninfected individuals after four days, and in most after 12 days cohabitation with infected flies. Iflavirus, but not cripavirus, was transmitted vertically, and surface-sterilised embryos contained high loads. Furthermore, high iflavirus loads in individual females resulted in high loads in their offspring. We demonstrated that viruses are highly prevalent in laboratory populations and that it is possible to establish and maintain uninfected fly lines for the assessment of virus transmission and host effects. This is important for pest management strategies such as the sterile insect technique which requires the mass-rearing of flies, as their fitness and performance may be affected by covert virus infections., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Background: Failure to comply with institutional and national standards for blood product administration poses patient safety risks. Monitoring adherence to blood product administration policy is important for maintaining patient safety and ensuring regulatory agency compliance., Study Design and Methods: This post-implementation quality improvement project evaluated an electronic paging system that was developed to facilitate perioperative blood product administration audits., Results: Of the 57 audits conducted, 55 (96.5%) audits were fully completed. Anesthesia providers verified the patient name and medical record number (MRN) on the patient's armband with the product ID tag in 36.5% (n = 20) of audits. Anesthesia providers ensured the name and MRN on the armband matched the compatibility label on the blood product in 23% (n = 23) of audits., Discussion: Ongoing surveillance of blood product administration practices is needed to monitor adherence to blood product administration policy, promote patient safety, and avoid imminent financial consequences from noncompliance to national regulatory standards., (© 2022 AABB.)

Maternally inherited bacterial endosymbionts that affect host fitness are common in nature. Some endosymbionts colonise host populations by reproductive manipulations (such as cytoplasmic incompatibility; CI) that increase the reproductive fitness of infected over uninfected females. Theory predicts that CI-inducing endosymbionts in haplodiploid hosts may also influence sex allocation, including in compatible crosses, however, empirical evidence for this is scarce. We examined the role of two common CI-inducing endosymbionts, Cardinium and Wolbachia, in the sex allocation of Pezothrips kellyanus, a haplodiploid thrips species with a split sex ratio. In this species, irrespective of infection status, some mated females are constrained to produce extremely male-biased broods, whereas other females produce extremely female-biased broods. We analysed brood sex ratio of females mated with males of the same infection status at two temperatures. We found that at 20 °C the frequency of constrained sex allocation in coinfected pairs was reduced by 27% when compared to uninfected pairs. However, at 25 °C the constrained sex allocation frequency increased and became similar between coinfected and uninfected pairs, resulting in more male-biased population sex ratios at the higher temperature. This temperature-dependent pattern occurred without changes in endosymbiont densities and compatibility. Our findings indicate that endosymbionts affect sex ratios of haplodiploid hosts beyond the commonly recognised reproductive manipulations by causing female-biased sex allocation in a temperature-dependent fashion. This may contribute to a higher transmission efficiency of CI-inducing endosymbionts and is consistent with previous models that predict that CI by itself is less efficient in driving endosymbiont invasions in haplodiploid hosts., (© 2022. The Author(s).)

Insect mitogenome organisation is highly conserved, yet, some insects, especially with parasitic life cycles, have rearranged mitogenomes. Furthermore, intraspecific mitochondrial diversity can be reduced by fitness-affecting bacterial endosymbionts like Wolbachia due to their maternal coinheritance with mitochondria. We have sequenced mitogenomes of the Wolbachia-infected endoparasitoid Dipterophagus daci (Strepsiptera: Halictophagidae) and four of its 22 known tephritid fruit fly host species using total genomic extracts of parasitised flies collected across > 700 km in Australia. This halictophagid mitogenome revealed extensive rearrangements relative to the four fly mitogenomes which exhibited the ancestral insect mitogenome pattern. Compared to the only four available other strepsipteran mitogenomes, the D. daci mitogenome had additional transpositions of one rRNA and two tRNA genes, and a single nucleotide frameshift deletion in nad5 requiring translational frameshifting or, alternatively, resulting in a large protein truncation. Dipterophagus daci displays an almost completely endoparasitic life cycle when compared to Strepsiptera that have maintained the ancestral state of free-living adults. Our results support the hypothesis that the transition to extreme endoparasitism evolved together with increased levels of mitogenome changes. Furthermore, intraspecific mitogenome diversity was substantially smaller in D. daci than the parasitised flies suggesting Wolbachia reduced mitochondrial diversity because of a role in D. daci fitness., (© 2022. The Author(s).)

Tephritid fruit flies are amongst the most devastating pests of horticulture, and Sterile Insect Technique (SIT) programs have been developed for their control. Their interactions with viruses are still mostly unexplored, yet, viruses may negatively affect tephritid health and performance in SIT programs, and, conversely, constitute potential biological control agents. Here we analysed ten transcriptome libraries obtained from laboratory populations of nine tephritid species from Australia (six species of Bactrocera, and Zeugodacus cucumis), Asia (Bactrocera dorsalis) and Europe (Ceratitis capitata). We detected new viral diversity, including near-complete (>99%) and partially complete (>80%) genomes of 34 putative viruses belonging to eight RNA virus families. On average, transcriptome libraries included 3.7 viruses, ranging from 0 (Z. cucumis) to 9 (B. dorsalis). Most viruses belonged to the Picornavirales, represented by fourteen Dicistroviridae (DV), nine Iflaviridae (IV) and two picorna-like viruses. Others were a virus from Rhabdoviridae (RV), one from Xinmoviridae (both Mononegavirales), several unclassified Negev- and toti-like viruses, and one from Metaviridae (Ortervirales). Using diagnostic PCR primers for four viruses found in the transcriptome of the Bactrocera tryoni strain bent wings (BtDV1, BtDV2, BtIV1, and BtRV1), we tested nine Australian laboratory populations of five species (B. tryoni, Bactrocera neohumeralis, Bactrocera jarvisi, Bactrocera cacuminata, C. capitata), and one field population each of B. tryoni, B. cacuminata and Dirioxa pornia. Viruses were present in most laboratory and field populations yet their incidence differed for each virus. Prevalence and co-occurrence of viruses in B. tryoni and B. cacuminata were higher in laboratory than field populations. This raises concerns about the potential accumulation of viruses and their potential health effects in laboratory and mass-rearing environments which might affect flies used in research and control programs such as SIT., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Wolbachia are widespread endosymbionts that affect arthropod reproduction and fitness. Mostly maternally inherited, Wolbachia are occasionally transferred horizontally. Previously, two Wolbachia strains were reported at low prevalence and titres across seven Australian tephritid species, possibly indicative of frequent horizontal transfer. Here, we performed whole-genome sequencing of field-caught Wolbachia-positive flies. Unexpectedly, we found complete mitogenomes of an endoparasitic strepsipteran, Dipterophagus daci, suggesting that Wolbachia in the flies are linked to concealed parasitization. We performed the first genetic characterization of D. daci and detected D. daci in Wolbachia-positive flies not visibly parasitized, and most but not all Wolbachia-negative flies were D. daci-negative, presumably reflecting polymorphism for the Wolbachia infections in D. daci. We dissected D. daci from stylopized flies and confirmed that Wolbachia infects D. daci, but also found Wolbachia in stylopized fly tissues, likely somatic, horizontally transferred, non-heritable infections. Furthermore, no Wolbachia cif and wmk genes were detected and very low mitogenomic variation in D. daci across its distribution. Therefore, Wolbachia may influence host fitness without reproductive manipulation. Our study of 13 tephritid species highlights that concealed early stages of strepsipteran parasitization led to the previous incorrect assignment of Wolbachia co-infections to tephritid species, obscuring ecological studies of this common endosymbiont and its horizontal transmission by parasitoids., (© 2021 Society for Applied Microbiology and John Wiley & Sons Ltd.)

Background: The endosymbiont Wolbachia can manipulate arthropod reproduction and invade host populations by inducing cytoplasmic incompatibility (CI). Some host species are coinfected with multiple Wolbachia strains which may have sequentially invaded host populations by expressing different types of modular CI factor (cif) genes. The tephritid fruit fly Rhagoletis cerasi is a model for CI and Wolbachia population dynamics. It is associated with at least four Wolbachia strains in various combinations, with demonstrated (wCer2, wCer4), predicted (wCer1) or unknown (wCer5) CI phenotypes., Results: We sequenced and assembled the draft genomes of the Wolbachia strains wCer1, wCer4 and wCer5, and compared these with the previously sequenced genome of wCer2 which currently invades R. cerasi populations. We found complete cif gene pairs in all strains: four pairs in wCer2 (three Type I; one Type V), two pairs in wCer1 (both Type I) and wCer4 (one Type I; one Type V), and one pair in wCer5 (Type IV). Wolbachia genome variant analyses across geographically and genetically distant host populations revealed the largest diversity of single nucleotide polymorphisms (SNPs) in wCer5, followed by wCer1 and then wCer2, indicative of their different lengths of host associations. Furthermore, mitogenome analyses of the Wolbachia genome-sequenced individuals in combination with SNP data from six European countries revealed polymorphic mitogenome sites that displayed reduced diversity in individuals infected with wCer2 compared to those without., Conclusions: Coinfections with Wolbachia are common in arthropods and affect options for Wolbachia-based management strategies of pest and vector species already infected by Wolbachia. Our analyses of Wolbachia genomes of a host naturally coinfected by several strains unravelled signatures of the evolutionary dynamics in both Wolbachia and host mitochondrial genomes as a consequence of repeated invasions. Invasion of already infected populations by new Wolbachia strains requires new sets of functionally different cif genes and thereby may select for a cumulative modularity of cif gene diversity in invading strains. Furthermore, we demonstrated at the mitogenomic scale that repeated CI-driven Wolbachia invasions of hosts result in reduced mitochondrial diversity and hitchhiking effects. Already resident Wolbachia strains may experience similar cytoplasmic hitchhiking effects caused by the invading Wolbachia strain., (© 2021. The Author(s).)

The draft genome sequence of a novel " Candidatus Liberibacter" species detected in an unidentified species of Zanthoxylum (Rutaceae) collected in Bhutan is reported. The total length is 1,408,989 bp with 1,169 coding sequences in 96 contigs, a GC content of 37.3%, and 76 to 77% average nucleotide identity with several other " Ca Liberibacter" species., (Copyright © 2020 Chambers et al.)

Background: Several plant-pathogenic bacteria are transmitted by insect vector species that often also act as hosts. In this interface, these bacteria encounter plant endophytic, insect endosymbiotic and other microbes. Here, we used high throughput sequencing to examine the bacterial communities of five different psyllids associated with citrus and related plants of Rutaceae in Bhutan: Diaphorina citri, Diaphorina communis, Cornopsylla rotundiconis, Cacopsylla heterogena and an unidentified Cacopsylla sp., Results: The microbiomes of the psyllids largely comprised their obligate P-endosymbiont 'Candidatus Carsonella ruddii', and one or two S-endosymbionts that are fixed and specific to each lineage. In addition, all contained Wolbachia strains; the Bhutanese accessions of D. citri were dominated by a Wolbachia strain first found in American isolates of D. citri, while D. communis accessions were dominated by the Wolbachia strain, wDi, first detected in D. citri from China. The S-endosymbionts from the five psyllids grouped with those from other psyllid taxa; all D. citri and D. communis individuals contained sequences matching 'Candidatus Profftella armatura' that has previously only been reported from other Diaphorina species, and the remaining psyllid species contained OTUs related to unclassified Enterobacteriaceae. The plant pathogenic 'Candidatus Liberibacter asiaticus' was found in D. citri but not in D. communis. Furthermore, an unidentified 'Candidatus Liberibacter sp.' occurred at low abundance in both Co. rotundiconis and the unidentified Cacopsylla sp. sampled from Zanthoxylum sp.; the status of this new liberibacter as a plant pathogen and its potential plant hosts are currently unknown. The bacterial communities of Co. rotundiconis also contained a range of OTUs with similarities to bacteria previously found in samples taken from various environmental sources., Conclusions: The bacterial microbiota detected in these Bhutanese psyllids support the trends that have been seen in previous studies: psyllids have microbiomes largely comprising their obligate P-endosymbiont and one or two S-endosymbionts. In addition, the association with plant pathogens has been demonstrated, with the detection of liberibacters in a known host, D. citri, and identification of a putative new species of liberibacter in Co. rotundiconis and Cacopsylla sp.

The application of Wolbachia in insect pest and vector control requires the establishment of genotypically stable host associations. The cytoplasmic incompatibility (CI) inducing Wolbachia strain wCer2 naturally occurs in the cherry fruit fly Rhagoletis cerasi as co-infection with other strains and was transferred to other fruit fly species by embryonic microinjections. We obtained wCer2 genome data from its native and three novel hosts, Drosophila simulans, Drosophila melanogaster, and Ceratitis capitata and assessed its genome stability, characteristics, and CI factor (cif) genes. De novo assembly was successful from Wolbachia cell-enriched singly infected D. simulans embryos, with minimal host and other bacterial genome traces. The low yield of Wolbachia sequence reads from total genomic extracts of one multiply infected R. cerasi pupa and one singly infected C. capitata adult limited de novo assemblies but was sufficient for comparative analyses. Across hosts wCer2 was stable in genome synteny and content. Polymorphic nucleotide sites were found in wCer2 of each host; however, only one nucleotide was different between R. cerasi and C. capitata, and none between replicated D. simulans lines. The wCer2 genome is highly similar to wAu (D. simulans), wMel (D. melanogaster), and wRec (Drosophila recens). In contrast to wMel and wRec (each with one cif gene pair) and wAu (without any cif genes), wCer2 has three pairs of Type I cif genes, and one Type V cifB gene without a cifA complement. This may explain previously reported CI patterns of wCer2, including incomplete rescue of its own CI modification in three novel host species., (© The Author(s) 2020. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.)

Background: Mass-rearing, domestication and gamma irradiation of tephritid fruit flies used in sterile insect technique (SIT) programmes can negatively impact fly quality and performance. Symbiotic bacteria supplied as probiotics to mass-reared fruit flies may help to overcome some of these issues. However, the effects of tephritid ontogeny, sex, diet and irradiation on their microbiota are not well known., Results: We have used next-generation sequencing to characterise the bacterial community composition and structure within Queensland fruit fly, Bactrocera tryoni (Froggatt), by generating 16S rRNA gene amplicon libraries derived from the guts of 58 individual teneral and mature, female and male, sterile and fertile adult flies reared on artificial larval diets in a laboratory or mass-rearing environment, and fed either a full adult diet (i.e. sugar and yeast hydrolysate) or a sugar only adult diet. Overall, the amplicon sequence read volume in tenerals was low and smaller than in mature adult flies. Operational taxonomic units (OTUs), belonging to the families Enterobacteriaceae (8 OTUs) and Acetobacteraceae (1 OTU) were most prevalent. Enterobacteriaceae dominated laboratory-reared tenerals from a colony fed a carrot-based larval diet, while Acetobacteraceae dominated mass-reared tenerals from a production facility colony fed a lucerne chaff based larval diet. As adult flies matured, Enterobacteriaceae became dominant irrespective of larval origin. The inclusion of yeast in the adult diet strengthened this shift away from Acetobacteraceae towards Enterobacteriaceae. Interestingly, irradiation increased 16S rRNA gene sequence read volume., Conclusions: Our findings suggest that bacterial populations in fruit flies experience significant bottlenecks during metamorphosis. Gut bacteria in teneral flies were less abundant and less diverse, and impacted by colony origin. In contrast, mature adult flies had selectively increased abundances for some gut bacteria, or acquired these bacteria from the adult diet and environment. Furthermore, irradiation augmented bacterial abundance in mature flies. This implies that either some gut bacteria were compensating for damage caused by irradiation or irradiated flies had lost their ability to regulate bacterial load. Our findings suggest that the adult stage prior to sexual maturity may be ideal to target for probiotic manipulation of fly microbiota to increase fly performance in SIT programmes.

Background: Obligate bacterial primary (P-) endosymbionts that are maternally inherited and codiverge with hosts are widespread across insect lineages with nutritionally restricted diets. Secondary (S-) endosymbionts are mostly facultative, but in some hosts, they complement P-endosymbiont function and therefore become obligate. Phylogenetic evidence exists for host switching and replacement of S-endosymbionts. The community dynamics that precede endosymbiont replacement and complementation have been little studied across host species, yet they are fundamental to the evolution of endosymbiosis., Results: We performed bacterial 16S rRNA gene amplicon sequencing of 25 psyllid species (Hemiptera, Psylloidea) across different developmental stages and ecological niches by focusing on the characterisation of the bacteria other than the universally present P-endosymbiont Carsonella (Gammaproteobacteria). Most species harboured only one dominant representative of diverse gammaproteobacterial S-endosymbionts that was consistently detected across all host individuals and populations (Arsenophonus in eight species, Sodalis or Sodalis-like bacteria in four species, unclassified Enterobacteriaceae in eight species). The identity of this dominant obligate S-endosymbiont varied across closely related host species. Unexpectedly, five psyllid species had two or three co-occurring endosymbiont species other than Carsonella within all host individuals, including a Rickettsiella-like bacterium (Gammaproteobacteria) in one psyllid species. Based on standard and quantitative PCR, all psyllids carried Carsonella, at higher titres than their dominant S-endosymbionts. Some psyllids also had Alphaproteobacteria (Lariskella, Rickettsia, Wolbachia) at varying prevalence. Incidence of other bacteria, including known plant pathogens, was low. Ecological niche of gall-forming, lerp-forming and free-living psyllid species did not impact endosymbiont communities. Two flush-feeding psyllid species had population-specific differences, and this was attributable to the higher endosymbiont diversity in native ranges and the absence of some endosymbionts in invasive ranges., Conclusions: Our data support the hypothesis of strict vertical transmission of minimal core communities of bacteria in psyllids. We also found evidence for S-endosymbiont replacement across closely related psyllid species. Multiple dominant S-endosymbionts present in some host species, including at low titre, constitute potential examples of incipient endosymbiont complementation or replacement. Our multiple comparisons of deep-sequenced minimal insect bacterial communities exposed the dynamics involved in shaping insect endosymbiosis.

Cardinium and Wolbachia are common maternally inherited reproductive parasites that can coinfect arthropods, yet interactions between both bacterial endosymbionts are rarely studied. For the first time, we report their independent expression of complete cytoplasmic incompatibility (CI) in a coinfected host, and CI in a species of the haplodiploid insect order Thysanoptera. In Pezothrips kellyanus, Cardinium-induced CI resulted in a combination of male development (MD) and embryonic female mortality (FM) of fertilized eggs. In contrast, Wolbachia-induced CI resulted in FM together with postembryonic mortality not previously reported as a CI outcome. Both endosymbionts appeared to not influence fecundity but virgins produced more offspring than mated females. In coinfected individuals, Wolbachia density was higher than Cardinium. Wolbachia removal did not impact Cardinium density, suggesting a lack of competition within hosts. Maternal transmission was complete for Wolbachia and high for Cardinium. Our data support theoretical predictions and empirical detection of high endosymbiont prevalence in field populations of the native range of this pest thrips. However, previous findings of more frequent loss of Wolbachia than Cardinium, particularly in field populations of the host's invasive range, suggest that genetic diversity or varying environmental factors between field populations also play a role in shaping host-endosymbiont dynamics., (© 2017 The Author(s). Evolution © 2017 The Society for the Study of Evolution.)

Fine particulate matter with less than 2.5 microns diameter (PM 2.5 ) generated by cattle in feedlots is an environmental pollutant and a potential human and animal health issue. The objective of this study was to determine if a feeding schedule affects cattle behaviors that promote PM 2.5 in a commercial feedlot. The study used 2813 crossbred steers housed in 14 adjacent pens at a large-scale commercial West Texas feedlot. Treatments were conventional feeding at 0700, 1000, and 1200 (CON) or feeding at 0700, 1000, and 1830 (ALT), the latter feeding time coincided with dusk. A mobile behavior lab was used to quantify behaviors of steers that were associated with generation of PM 2.5 (e.g., fighting, mounting of peers, and increased locomotion). PM 2.5 samplers measured respirable particles with a mass median diameter ≤2.5 μm (PM 2.5 ) every 15 min over a period of 7 d in April and May. Simultaneously, the ambient temperature, humidity, wind speed and direction, precipitation, air pressure, and solar radiation were measured with a weather station. Elevated downwind PM 2.5 concentrations were measured at dusk, when cattle that were fed according to the ALT vs. the CON feeding schedule, demonstrated less PM 2.5 -generating behaviors ( p < 0.05). At dusk, steers on ALT vs. CON feeding schedules ate or were waiting to eat (standing in second row behind feeding cattle) at much greater rates ( p < 0.05). Upwind PM 2.5 concentrations were similar between the treatments. Downwind PM 2.5 concentrations averaged over 24 h were lower from ALT compared with CON pens (0.072 vs. 0.115 mg/m³, p < 0.01). However, dry matter intake (DMI) was less ( p < 0.05), and average daily gain (ADG) tended to be less ( p < 0.1) in cattle that were fed according to the ALT vs. the CON feeding schedules, whereas feed efficiency (aka gain to feed, G:F) was not affected. Although ALT feeding may pose a challenge in feed delivery and labor scheduling, cattle exhibited fewer PM 2.5 -generating behaviors and reduced generation of PM 2.5 when feed delivery times matched the natural desires of cattle to eat in a crepuscular pattern.

Coevolution between insects and bacterial endosymbionts contributes to the success of many insect lineages. For the first time, we tested for phylogenetic codivergence across multiple taxonomic scales, from within genera to superfamily between 36 psyllid species of seven recognised families (Hemiptera: Psylloidea), their exclusive primary endosymbiont Carsonella and more diverse secondary endosymbionts (S-endosymbionts). Within Aphalaridae, we found that Carsonella and S-endosymbionts were fixed in one Glycaspis and 12 Cardiaspina populations. The dominant S-endosymbiont was Arsenophonus, while Sodalis was detected in one Cardiaspina species. We demonstrated vertical transmission for Carsonella and Arsenophonus in three Cardiaspina species. We found strong support for strict cospeciation and validated the informative content of Carsonella as extended host genome for inference of psyllid relationships. However, S-endosymbiont and host phylogenies were incongruent, and displayed signs of host switching and endosymbiont replacement. The high incidence of Arsenophonus in psyllids and other plant sap-feeding Hemiptera may be due to repeated host switching within this group. In two psyllid lineages, Arsenophonus and Sodalis genes exhibited accelerated evolutionary rates and AT-biases characteristic of long-term host associations. Together with strict vertical transmission and 100% prevalence within host populations, our results suggest an obligate, and not facultative, symbiosis between psyllids and some S-endosymbionts., (© 2016 Society for Applied Microbiology and John Wiley & Sons Ltd.)

Background: Maternally inherited Wolbachia bacteria infect many insect species. They can also be transferred horizontally into uninfected host lineages. A Wolbachia spillover from an infected source population must occur prior to the establishment of heritable infections, but this spillover may be transient. In a previous study of tephritid fruit fly species of tropical Australia we detected a high incidence of identical Wolbachia strains in several species as well as Wolbachia pseudogenes. Here, we have investigated this further by analysing field specimens of 24 species collected along a 3,000 km climate gradient of eastern Australia., Results: Wolbachia sequences were detected in individuals of nine of the 24 (37 %) species. Seven (29 %) species displayed four distinct Wolbachia strains based on characterisation of full multi locus sequencing (MLST) profiles; the strains occurred as single and double infections in a small number of individuals (2-17 %). For the two remaining species all individuals had incomplete MLST profiles and Wolbachia pseudogenes that may be indicative of lateral gene transfer into host genomes. The detection of Wolbachia was restricted to northern Australia, including in five species that only occur in the tropics. Within the more widely distributed Bactrocera tryoni and Bactrocera neohumeralis, Wolbachia also only occurred in the north, and was not linked to any particular mitochondrial haplotypes., Conclusions: The presence of Wolbachia pseudogenes at high prevalence in two species in absence of complete MLST profiles may represent footprints of historic infections that have been lost. The detection of identical low prevalence strains in a small number of individuals of seven species may question their role as reproductive manipulator and their vertical inheritance. Instead, the findings may be indicative of transient infections that result from spillover events from a yet unknown source. These spillover events appear to be restricted to northern Australia, without proliferation in host lineages further south. Our study highlights that tropical fruit fly communities contain Wolbachia pseudogenes and may be exposed to frequent horizontal Wolbachia transfer. It also emphasises that global estimates of Wolbachia frequencies may need to consider lateral gene transfer and Wolbachia spillover that may be regionally restricted, transient and not inherited.

Tephritid fruit fly species display a diversity of host plant specialisation on a scale from monophagy to polyphagy. Furthermore, while some species prefer ripening fruit, a few are restricted to damaged or rotting fruit. Such a diversity of host plant use may be reflected in the microbial symbiont diversity of tephritids and their grade of dependency on their microbiomes. Here, we investigated the microbiome of six tephritid species from three genera, including species that are polyphagous pests (Bactrocera tryoni, Bactrocera neohumeralis, Bactrocera jarvisi, Ceratitis capitata) and a monophagous specialist (Bactrocera cacuminata). These were compared with the microbiome of a non-pestiferous but polyphagous tephritid species that is restricted to damaged or rotting fruit (Dirioxa pornia). The bacterial community associated with whole fruit flies was analysed by 16S ribosomal DNA (rDNA) amplicon pyrosequencing to detect potential drivers of taxonomic composition. Overall, the dominant bacterial families were Enterobacteriaceae and Acetobacteraceae (both Proteobacteria), and Streptococcaceae and Enterococcaceae (both Firmicutes). Comparisons across species and genera found different microbial composition in the three tephritid genera, but limited consistent differentiation between Bactrocera species. Within Bactrocera species, differentiation of microbial composition seemed to be influenced by the environment, possibly including their diets; beyond this, tephritid species identity or ecology also had an effect. The microbiome of D. pornia was most distinct from the other five species, which may be due to its ecologically different niche of rotting or damaged fruit, as opposed to ripening fruit favoured by the other species. Our study is the first amplicon pyrosequencing study to compare the microbiomes of tephritid species and thus delivers important information about the turnover of microbial diversity within and between fruit fly species and their potential application in pest management strategies.

Wolbachia are endosymbiotic bacteria that infect 40-65% of arthropod species. They are primarily maternally inherited with occasional horizontal transmission for which limited direct ecological evidence exists. We detected Wolbachia in 8 out of 24 Australian tephritid species. Here, we have used multilocus sequence typing (MLST) to further characterize these Wolbachia strains, plus a novel quantitative polymerase chain reaction method for allele assignment in multiple infections. Based on five MLST loci and the Wolbachia surface protein gene (wsp), five Bactrocera and one Dacus species harboured two identical strains as double infections; furthermore, Bactrocera neohumeralis harboured both of these as single or double infections, and sibling species B. tryoni harboured one. Two Bactrocera species contained Wolbachia pseudogenes, potentially within the fruit fly genomes. A fruit fly parasitoid, Fopius arisanus shared identical alleles with two Wolbachia strains detected in one B. frauenfeldi individual. We report an unprecedented high incidence of four shared Wolbachia strains in eight host species from two trophic levels. This suggests frequent exposure to Wolbachia in this tropical tephritid community that shares host plant and parasitoid species, and also includes species that hybridize. Such insect communities may act as horizontal transmission platforms that contribute to the ubiquity of the otherwise maternally inherited Wolbachia., (© 2014 Society for Applied Microbiology and John Wiley & Sons Ltd.)

In tephritids, the sex-determination pathway follows the sex-specific splicing of transformer (tra) mRNA, and the cooperation of tra and transformer-2 (tra-2) to effect the sex-specific splicing of doublesex (dsx), the genetic double-switch responsible for male or female somatic development. The Dominant Male Determiner (M) is the primary signal that controls this pathway. M, as yet uncharacterized, is Y-chromosome linked, expressed in the zygote and directly or indirectly diminishes active TRA protein in male embryos. Here we first demonstrated the high conservation of tra, tra-2 and dsx in two Australian tephritids, Bactrocera tryoni and Bactrocera jarvisi. We then used quantitative reverse transcription PCR on single, sexed embryos to examine expression of the key sex-determination genes during early embryogenesis. Individual embryos were sexed using molecular markers located on the B. jarvisi Y-chromosome that was also introgressed into a B. tryoni line. In B. jarvisi, sex-specific expression of tra transcripts occurred between 3 to 6 h after egg laying, and the dsx isoform was established by 7 h. These milestones were delayed in B. tryoni lines. The results provide a time frame for transcriptomic analyses to identify M and its direct targets, plus information on genes that may be targeted for the development of male-only lines for pest management., (© 2014 The Royal Entomological Society.)

Background: Developing embryos are provided with maternal RNA transcripts and proteins, but transcription from the zygotic nuclei must be activated to control continuing embryonic development. Transcripts are generated at different stages of early development, and those involved in sex determination and cellularisation are some of the earliest to be activated. The male sex in tephritid fruit flies is determined by the presence of a Y chromosome, and it is believed that a transcript from the Y-chromosome sets in motion a cascade that determines male development, as part of the greater maternal to zygotic transition (MTZ). Here we investigate the poly(A+) transcriptome in early male and female embryos of the horticultural pest Bactrocera jarvisi (Diptera: Tephritidae)., Results: Bactrocera jarvisi embryos were collected over two pre-blastoderm time periods, 2-3h and 3-5h after egg laying. Embryos were individually sexed using a Y-chromosome marker, allowing the sex-specific poly(A+) transcriptome of single-sex embryo pools to be deep-sequenced and assembled de novo. Transcripts for sixteen sex-determination and two cellularisation gene homologues of Drosophila melanogaster (Diptera: Drosophilidae) were identified in early embryos of B. jarvisi, including transcripts highly upregulated prior to cellularisation. No strong candidates for transcripts derived solely from the Y chromosome were recovered from the poly(A+) fraction., Conclusions: Bactrocera jarvisi provides an excellent model for embryonic studies due to available Y-chromosome markers and the compact time frame for zygotic transcription and the sex-determined state. Our data contribute fundamental information to sex-determination research, and provide candidates for the sourcing of gene promoters for transgenic pest-management strategies of tephritid fruit flies.

Among Australian endemic tephritid fruit flies, the sibling species Bactrocera tryoni and Bactrocera neohumeralis have been serious horticultural pests since the introduction of horticulture in the nineteenth century. More recently, Bactrocera jarvisi has also been declared a pest in northern Australia. After several decades of genetic research there is now a range of classical and molecular genetic tools that can be used to develop improved Sterile Insect Technique (SIT) strains for control of these pests. Four-way crossing strategies have the potential to overcome the problem of inbreeding in mass-reared strains of B. tryoni. The ability to produce hybrids between B. tryoni and the other two species in the laboratory has proved useful for the development of genetically marked strains. The identification of Y-chromosome markers in B. jarvisi means that male and female embryos can be distinguished in any strain that carries a B. jarvisi Y chromosome. This has enabled the study of homologues of the sex-determination genes during development of B jarvisi and B. tryoni, which is necessary for the generation of genetic-sexing strains. Germ-line transformation has been established and a draft genome sequence for B. tryoni released. Transcriptomes from various species, tissues and developmental stages, to aid in identification of manipulation targets for improving SIT, have been assembled and are in the pipeline. Broad analyses of the microbiome have revealed a metagenome that is highly variable within and across species and defined by the environment. More specific analyses detected Wolbachia at low prevalence in the tropics but absent in temperate regions, suggesting a possible role for this endosymbiont in future control strategies.

We report the heritable germ-line transformation of the Queensland fruit fly, Bactrocera tryoni, using a piggyBac vector marked with either the fluorescent protein DsRed or EGFP. A transformation frequency of 5-10% was obtained. Inheritance of the transgenes has remained stable over more than 15 generations despite the presence of endogenous piggyBac sequences in the B. tryoni genome. The sequence of insertion sites shows the usual canonical pattern of piggyBac integraton into TTAA target sites. An investigation of endogenous piggyBac elements in the B. tryoni genome reveals the presence of sequences almost identical to those reported recently for the B. dorsalis complex of fruit flies and two noctuid moths, suggesting a common origin of piggyBac sequences in these species. The availability of transformation protocols for B. tryoni has the potential to deliver improvements in the performance of the Sterile Insect Technique for this pest species.

Bactrocera tryoni (Froggatt) (Diptera: Tephritidae) or "Qfly," is the most serious horticultural pest in Australia, with a bioclimatic range that extends from the tropical north to the temperate south. Various Australian horticultural exports depend on certification that they originated from B. tryoni-free areas. To eliminate, rather than suppress, B. tryoni in production areas, a sterile insect technique (SIT) campaign directed at B. tryoni has been in operation in southeastern Australia since 1997. Like many other SIT programs around the world, the B. tryoni SIT program relies on fluorescent dust to mark the sterile insects. However, fluorescent dust marking does not provide 100% accuracy in the identification of sterile insects, as required where the aim is to declare regions completely free of fruit fly. Here, we show that novel mitochondrial markers can be introduced into a strain of B. tryoni by interspecies hybridization between B. tryoni and a related but well-differentiated species, Bactrocera jarvisi (Tryon), followed by backcrossing of the hybrid strain with the parental B. tryoni strain. These novel markers do not affect the viability of the strain as measured by pupation and eclosion rates. A simple polymerase chain reaction-based test is described that distinguishes the marked B. tryoni from wild B. tryoni. As required in practice, the test was shown to work reliably on DNA extracted from dead flies that had remained in field traps for up to two weeks.

The objectives of this study were to determine the effects of shading wallows during the summer months on lactating sow behavior, performance, and physiology. A total of 128 sows were used during warm weather (May to August 2001) to determine the effects of unshaded (control; n = 8) vs. shaded (SH; n = 8) wallows. Sows ranged over 6 parities and were fed a completely balanced sorghum-based diet. Behavioral data were collected by 15-min scan samples over a 24-h period/wk for a total of 16 wk. All sows were observed twice when litter age was 5 and 15 d, respectively. Respiration rates (breaths/min) were collected on 50 sows (control, n = 25; SH, n = 25) over an 8-wk period when the maximum temperature exceeded 32 degrees C. Ten milliliters of clotted blood and 20 mL of whole blood were obtained by jugular puncture from each sow on the day of weaning to determine total white blood cells, acute phase proteins, packed-cell volume, and chemotaxis and chemokinesis. Descriptive water temperature profiles were measured by using data loggers positioned at 3 levels per wallow: surface water, shallow mud, and deep mud. Behavioral, postural, location, performance, and physiological measurements did not differ (P > 0.05) among wallow treatments. Regardless of treatment, sows spent approximately 82% of their total time budget inside the farrowing hut and only approximately 7% of their total time budget in the wallow. A total of 428 piglets died, 219 in the control treatment and 209 in the SH treatment. The majority of piglets in both treatments died of crushing within the first 72 h after parturition, and most of the piglets had suckled. Shade kept the shallow water profile cooler during the hotter afternoon temperatures compared with the control wallows. In SH for both the shallow and deep mud profiles, temperatures were consistent throughout the day. In conclusion, sows spent a large percentage of their daily time budget inside the farrowing hut and spent only brief episodes in the wallow. Shading the wallow did not result in increased wallow use time or improvements in sow physiology and overall performance.

The objective of this study was to more fully define the surgical stress response to dehorning by heat cauterization in dairy calves by measuring behavioral, hormonal, inflammatory, and immunological markers of stress and to determine whether a nerve block of the surgical site with a concentrated solution of lidocaine (5%) reduces the degree of stress. Thirty-two 10- to 12-wk-old female Holstein calves were randomly allotted to 1 of 4 treatments: 5% lidocaine followed by dehorning, 2% lidocaine followed by dehorning, saline followed by dehorning, or 5% lidocaine followed by sham dehorning. Plasma cortisol concentration was measured in blood samples collected via a jugular catheter at -0.5, 0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 6, 9, 12, 24, 48, and 72 h. Various other blood constituents were measured in samples collected at -0.5, 12, 24, 48, and 72 h. Feeding, drinking, scratching, grooming, rubbing, licking, and inactivity behaviors were observed in the standing and recumbent positions using a 10-min scan sampling method analyzed on a time period and daily basis for 72 h following the dehorning procedure. The frequency of vocalization, kicking, and lying in the chute during the dehorning procedure were also assessed. The overall plasma cortisol concentrations were higher in calves subjected to dehorning than in control calves. Compared with the control group, the saline-treated calves had a higher cortisol concentration at 30 and 60 min postdehorning. Plasma cortisol concentrations were higher in all groups at 30 min postdehorning than at other sampling times. The percentage of circulating neutrophils and the neutrophil:lymphocyte ratio were increased in the saline and 2% lidocaine group. Total plasma protein, fibrinogen, and alpha1-acid glycoprotein concentrations were similar among treatments. The behavioral response to dehorning, as manifested by kicking while in the chute, was greater in the saline and 2% lidocaine group than in the control or 5% lidocaine treatment groups. In the postdehorning period, the percentage of time calves spent performing various maintenance behaviors did not differ among treatments. Thus, injection of 5% lidocaine may not provide any added comfort after the dehorning but may decrease the overall stress response during the procedure.

"Segregated early weaning" (SEW) of pigs reduces exposure to pathogenic bacteria, but upon arrival at grower facilities pigs may be co-mingled regardless of farm of origin. The present study was designed to examine the effect of mixing (social) stress on populations of Salmonella enterica Typhimurium in SEW pigs. Piglets (7 days old; n = 28 in each of 2 replicates) were separated into 2 treatments (control and mixed groups) of 2 pens per treatment (7 piglets/pen). One (n = 1) "seeder" pig/pen was inoculated with 10(9) CFU of S. Typhimurium. Each seeder was placed with non-inoculated "contact" piglets (n = 6). A"contact" piglet was swapped each day between the "mixed" pens for 5 days; pigs in control pens were not exchanged. On day 5, the incidence of fecal Salmonella shedding was higher in the mixed contact pigs (P < 0.05). Rectal Salmonella and cecal coliform populations in mixed pigs were significantly (P < 0.05) greater than in control pigs but cecal Salmonella populations were not different. Mixed pigs were more susceptible to tissue invasiveness (i.e., Salmonella-positive tonsils and lymph nodes) than control pigs. These results indicate that social stress of weaned pigs may increase susceptibility to and/or fecal shedding of Salmonella. Food-borne Salmonella infections in the United States are estimated to cost the economy dollar 2.4 billion annually (ERS/USDA, 2001). Approximately 6-9% of human salmonellosis is associated with the consumption of pork products (Frenzen et al., 1999). Salmonella is relatively common on swine farms and has been isolated from all stages of the pork production chain (Davies et al., 1999; Fedorka-Cray et al., 1997b; Rostagno et al., 2003). Salmonella is a threat to the pork industry not only from a food-safety perspective as a public health concern, but some Salmonella serotypes can cause clinical illnesses in swine, negatively impacting production efficiency and profitability (Schwartz, 1991).

Swine can harbor Salmonella in their gastrointestinal tracts. It has been estimated that up to 48% of the U.S. swine herd may carry Salmonella. Housing sows in farrowing stalls has become controversial due to animal welfare-based criticisms. An alternative production system is to keep sows outdoors on pasture with access to individual farrowing huts. This study was designed to determine the effects of two production systems on indicator bacteria and Salmonella of sows housed indoors in farrowing stalls (n = 52) compared to sows housed outdoors (n = 52) in English style huts. Each farrowing radial contained one wallow, from which mud (n = 290) and water (n = 290) samples were collected weekly. All samples were analyzed for generic E. coli, coliforms and Salmonella. No differences (p > 0.05) were detected in Salmonella, generic E. coli and coliform populations between indoor farrowing stalls and outdoor farrowing huts. However, all 8 outdoor wallows contained Salmonella spp. at some point during the study (n = 49 Salmonella isolates). Salmonella genotypes persisted within some wallows for >5 months, and genetically indistinguishable Salmonella isolates were found in multiple wallows. Salmonella isolated from outdoor sow feces were genetically indistinguishable by PFGE from Salmonella isolated from wallows (n = 33) throughout the study, indicating that pathogenic bacteria were cycling between swine and their environment. In conclusion, the role of wallows in disseminating Salmonella within an outdoor swine herd appears to be significant.

We have developed a technique for expanding primary cultures of human tracheal epithelium while minimizing loss of differentiated structure and function. Cells were seeded at 2 x 10(4) cells/cm2 into T75 flasks and trypsinized when approximately 80% confluent. The dispersed cells were then passaged at the same plating density into further T75 flasks or seeded at 5 x 10(5) cells/cm2 on porous-bottomed inserts and maintained with an air-interface. Differentiation of cells on inserts was assessed from transepithelial electrical resistance (an index of tight junction formation), short-circuit current (an index of transepithelial salt transport), cell numbers, total cell protein, and histology. Unpassaged cells (P0) and cells passaged once (P1) took about a week to become 80% confluent on T75 flasks, with 10-fold and 5-fold increases in cell numbers, respectively. Confluence was achieved in approximately 3 days following plating to inserts. Functionally and structurally, P1 and P2 cells (cells passaged twice) were little different from P0 cells. Thus, within a little over 2 weeks, the numbers of confluent cell sheets can be increased 50-fold with minimal change in function. However, there was a marked decline in differentiation by cells passaged three times (P3), and not all cell preparations could be taken to P4 (cells passaged four times).

Heat stress and dusty conditions are common challenges for cattle during the summer, and a typical method of alleviating these problems involves sprinkling cattle and pens with water. The effect of sprinkling water on the incidence of zoonotic pathogens has not been previously studied. Four pens of heifers (n = 41) were cooled using sprinklers, and four pens (n = 43) served as controls. Heifers were crossbred Charolais, with white and red hair coats. Sprinkling was initiated when cattle were on full concentrate feed (July). Fecal samples, hide swipes, and BW were collected on d 0, 28, 63, 95, and 98. Average daily gain, DMI, and G:F were calculated, and carcass traits were collected 36 h after processing. Performance data were analyzed as a randomized complete block design, and zoonotic pathogen data were analyzed using chi2 analysis. Sprinkling tended (P = 0.054) to increase the incidence of fecal Salmonella spp. populations on d 98, but simultaneously tended to decrease (P = 0.058) the Escherichia coli O157:H7 incidence on hides on d 98. The most prevalent Salmonella serovars in this study were Kentucky, Muenster, Meleagridis, and Cerro. Performance measures and carcass traits did not differ between treatments (P > 0.10). Under our conditions, sprinkling cattle with water did not affect the incidence of zoonotic pathogens in feces or on hides.

Plasma cortisol, porcine corticosteroid-binding globulin (pCBG), hepatic CBG expression, and other physiological and behavioral measures of stress were studied in pigs in response to elevated temperature in conjunction with establishing a social hierarchy. Twenty-four crossbred pigs were weaned at 25 d of age (three or six pigs from six sows) and housed in littermate groups at 23 +/- 2 degrees C. At 57 d of age (d 0), animals were weighed and placed under general anesthesia for collection of blood (10 mL) and liver (approximately 100 mg) samples. On d 1, three unacquainted pigs of similar BW (23 +/- 1 kg) from different litters were allotted to each of eight nursery pens within two environmentally controlled rooms (12 pigs per room). From d 1 to 7, one room was maintained at 23 +/- 2 degrees C (CON) and the other at 33 +/- 2 degrees C (HEAT). Both rooms were kept at 23 +/- 2 degrees C from d 8 to 14. Animals were videotaped for 72 h beginning on d 1 and 8 to document behavioral changes in response to room temperature. The social hierarchy of pigs within each pen was based on fight activity recorded on d 1 to 3. Blood and liver tissue were collected again on d 7 and 14. The ADG for HEAT pigs increased (P < 0.05) over d 8 to 14 compared with d 1 to 7. In contrast to CON pigs, HEAT pigs displayed increased (P < 0.01) drinking but decreased feeding and lying in contact with other pigs from d 1 to 3, and similar drinking and feeding but increased (P < 0.01) lying with contact behaviors from d 8 to 10. With the exception of subordinate pigs exhibiting less (P < 0.05) frequent standing/walking behavior than the dominant or intermediate pigs on d 1 to 3, frequency of behaviors for both recorded time periods did not differ among pigs due to social status, regardless of treatment. The concentration of plasma haptoglobin in HEAT pigs on d 7 compared with d 0 increased (467 vs. 763 mg/L; P < 0.05), whereas cortisol and pCBG decreased (274 vs. 235 nmol/L and 11.4 vs. 9.9 mg/L, respectively; P < 0.05) as a result of treatment. The free cortisol index (total cortisol/pCBG) was greater (P < 0.05) in HEAT pigs on d 14 than on d 0 or 7. Hepatic CBG mRNA level was not affected by treatment. On d 14, HEAT pigs had plasma cortisol, pCBG, and haptoglobin concentrations similar to those of CON pigs. These results indicate that measured behavioral and physiological responses were not related to social status, and decreased circulating levels of cortisol and pCBG in pigs following a 7-d exposure to elevated temperature may not be determined by hepatic CBG mRNA expression.

The period gene is important for the generation and maintenance of biological rhythms. It served as an ideal candidate for the investigation of the mating time isolation between two sibling Queensland fruit fly species, Bactrocera tryoni and Bactrocera neohumeralis. We have isolated the homologues of the period gene in the two species, and show that their putative amino acid sequences are identical. No length polymorphism was detected in the Thr-Gly repeat region. per mRNA expression, assayed in light-dark diurnal conditions, displayed circadian oscillation in both the head and abdomen of B. tryoni and B. neohumeralis, with the same cycling phase. An alternatively spliced intron was identified in the 3' untranslated region. The effect of temperature on the splicing and mRNA expression was examined.

A 10-year-old, lethargic, potbellied pig presented with signs of abdominal discomfort and a palpable abdominal mass. Laparotomy revealed a 20 cm diameter mass on the spleen and smaller masses on the omentum and liver. After euthanasia and histologic examination of the hepatic mass, the diagnosis was hepatocellular carcinoma.

One hundred twenty 8-wk-old barrows (20.3 +/- 2.0 kg BW) were used to examine the effect of split marketing on selected behavioral, physiological and performance parameters. Pigs were assigned by weight in a randomized complete block design to one of three treatments: SM (split-marketed), six pigs/pen (1.83 m2/pig); C (control), six pigs/pen (1.83 m2/pig); or MC (modified control), three pigs/pen (3.66 m2/pig). The heaviest half of SM animals were removed 1 wk prior to marketing penmates. Control and MC animals remained in their respective groups until marketing. Animals were videotaped during the first 72 h of the study (INITIAL), 72 h prior to (PRE), and following the removal (POST) of pigs in the SM treatment to quantify maintenance behaviors and to identify socially dominant, intermediate, and submissive pigs. A blood sample was collected from each animal upon completion of INITIAL, PRE, and POST time periods to determine neutrophil:lymphocyte ratio and plasma haptoglobin, cortisol, and corticosteroid-binding globulin (CBG) levels. Animals were weighed and feed disappearance was calculated biweekly. Tenth-rib backfat and area of the longissimus muscle at marketing were ultrasonically evaluated on all animals. Regardless of treatment, animals were more (P < 0.01) active (eating, standing/walking, fighting) at INITIAL than at PRE or POST times. Frequency and duration of fights per pen were less (P < 0.01) in MC than in C or SM pigs for all periods observed. Neutrophil:lymphocyte ratio, plasma haptoglobin, and CBG levels were greater (P < 0.01) during the INITIAL period than during the PRE or POST periods but did not differ between treatments. No treatment or time differences were detected in plasma cortisol levels. The MC pigs exhibited greater (P < 0.01) ADFI with poorer feed efficiency compared to C or SM pigs up to split marketing. During the POST period, both MC and SM pigs had greater (P < 0.01) ADFI with poorer (P < 0.01) feed efficiency than C pigs. The ADG was not different among animals as a result of treatment. There were no treatment differences for any of the carcass measurements. Significant differences in performance between the treatment groups could not be attributed to any physiological or behavioral measures reported here.

One hundred twelve crossbred feedlot heifers were used in two experiments to assess the impact of heat stress and its relief by shade and(or) water misting on behavior, physiology, performance, and carcass traits. Treatments were 1) no shading or misting (CONT); 2) only misting (MIST); 3) only shading (SHADE); and 4) shading plus misting (SHMI). Head in the feed bunk, head in or above the waterer, walking, standing, and lying behaviors were observed. Rectal temperature, respiration rate, and carcass traits were measured, as well as DMI, ADG, and feed:gain. Dietary NEm and NEg concentrations were calculated from performance data. In Exp. 1, (32 heifers; average BW 288 kg) the CONT heifers spent more time lying down than all others (P < 0.01). In addition, CONT heifers spent less time (P < 0.01) standing than SHADE and MIST heifers. Misting decreased (P < 0.01) rectal temperature and MIST as well as SHADE lowered (P < 0.05) respiration rates. In Exp. 2 (80 heifers; average BW = 336 kg), lying and walking behaviors did not differ among treatments, but shade increased (P < 0.01) standing behavior in heifers. The MIST cattle performed less (P < 0.05) head-above-water behavior than unmisted cattle. Rectal temperatures did not differ among treatments, but respiration rate was lower in shaded than in unshaded heifers (P < 0.05). Shaded compared with unshaded heifers had greater DMI (9.46 vs 8.80 +/- 0.14 kg/d, P < 0.01) and ADG (1.6 vs 1.41 +/- 0.1 kg/d, P < 0.01). Heifers provided with shade reached their target BW 20 d earlier than the unshaded heifers and differed in final BW (547 vs 520 +/- 6 kg, P < 0.01). Feed:gain and calculated NEg and NEm concentrations did not differ among treatments, and carcass traits were generally similar among treatments. In conclusion, cattle without shade had a physiological and behavioral stress response to heat that negatively affected productivity. Providing shade for beef cattle was a suitable solution to decrease heat stress and to lower the negative effects of heat on performance, whereas misting was largely ineffective.

One hundred twenty-four prepuberal crossbred pigs were used in a series of behavioral bioassays to determine the minimum dose of androstenone (5 alpha-androst-16-en-3-one) that would reduce the level of agonistic behavior among dyads of newly regrouped pigs. Randomly selected females and castrated males were used in 21-h videotaped observation periods. In Exp. 1, isopropyl alcohol was tested against no aerosol to determine if the vehicle (isopropyl alcohol) influenced agonistic behavior. Level of submissive and aggressive behaviors were similar (P greater than .10) between treatments. In Exp. 2, vehicle or androstenone in vehicle was sprayed on the snout and head of both pigs at the start of the encounter. Four bioassays were performed with four levels (.05, .5, 5 and 50 micrograms/pig) of androstenone dissolved in isopropyl alcohol. Sprayed isopropyl alcohol served as a control. At concentrations of .5 and 5 micrograms/pig, androstenone reduced aggressive behavior (P less than .05). Androstenone had no consistent effect on submissive behavior. In Exp. 3, androstenone was sprayed on pigs at the start of the encounter and again at 30, 60 and 90 min after pairs of pigs were mixed. Repeated application of this androgen resulted in levels of agonistic behavior similar to those recorded when nothing was applied (P greater than .10). A single application of as little as .5 micrograms androstenone per pig reduced aggressive behavior among prepuberal pigs and, therefore, may be a way of reducing fighting among newly regrouped prepuberal pigs.

Three experiments were conducted to evaluate sow and piglet productivity under extended photoperiod. In Exp. 1, 98 crossbred, lactating sows were housed in one of four treatments: thermoneutral air temperature (23.6 degrees C) in either (h of light:dark) 1:23 or 16:8 photoperiods, or heat stress (30.4 degrees C) in either 1:23 or 16:8 photoperiods. Heat stress reduced (P less than .05) sow feed intake, piglet mortality and piglet weaning weight and increased (P less than .01) sow lactation weight loss. Number of pigs weaned per litter was increased (P less than .01) when sows were heat-stressed. Extended photoperiod reduced (P greater than .05) time for sows to rebreed postweaning by .4 d. The interaction between air temperature and photoperiod was significant only for sow lactation weight loss. Heat stress increased sow lactation weight loss, but this effect was more severe in the 1:23 than in the 16:8 photoperiod. Experiments 2 and 3 examined the effects of 1:23 or 16:8 photoperiods on nursery pig performance when pigs were weaned from sows experiencing 1:23 (Exp. 3) or 16:8 (Exp. 2) photoperiods. In both nursery studies, photoperiod did not influence (P greater than .10) postweaning pig mortality, feed intake, weight gain or gain:feed ratio. In conclusion, extended photoperiod reduced days to return to estrus and reduced sow lactation weight loss, especially during heat stress. No benefits in preweaning or postweaning piglet weight or survival were observed by use of extended photoperiod.