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Recently, Wolbachia infection has been described in leishmaniasis vector sandflies. This endosymbiont bacterium is present in 60% of insects, and has been suggested as a mechanism of biological control of vector insects, because it causes a series of changes in the invertebrate host. In addition, recent studies have shown that this bacterium can prevent the development of parasites in vector insects. In this context, the present study aims to molecularly characterize the circulating strain of this bacterium in sandflies in the State of Bahia, Brazil, as well as the natural infection rate of Leishmania sp., and to evaluate the coinfection between Wolbachia and Leishmania. Seven hundred and forty-five (745) specimens of sandflies were collected in nine municipalities of Bahia, belonging to two species, Lutzomyia longipalpis (Lutz and Neiva, 1912) and Nyssomyia whitmani (Antunes and Coutinho, 1939). The results confirm infection by the protozoan Leishmania infantum and Wolbachia in both species collected. The identified strain of Wolbachia in sandflies was wStv MI, known to lead to a phenotype of cytoplasmic incompatibility in vector insects.

Benzimidazole (BZ) resistance of Haemonchus contortus has been associated with single nucleotide polymorphisms (SNPs) in codons 200 (F200Y) and 167 (F167Y) and, to a lesser extent, in codon E198A, of the β-tubulin isotype 1 gene. The present study was undertaken to survey the status of BZ resistance in naturally infected goats in smallholder farms in southern Mozambique by real-time PCR (qPCR) using TaqMan® assays. H. contortus-infective larvae (L3; n = 432) from 12 populations were individually genotyped for F200Y and F167Y SNPs to detect BZ resistance. For the F200Y SNP, the results revealed an overall mean percentages of 18.8% homozygous resistant (RR), 47.8% homozygous susceptible (SS) and 33.4% heterozygous (RS) H. contortus. For the F167Y SNP, the overall mean percentages were 1.6% RR, 94.9% SS and 3.5% RS. The percentage of resistant alleles (%R) for the F200Y and F167Y SNPs was 35.7 and 3.4%, respectively. Genotype combinations of the two mutations indicate resistant percentages ranging from 0.0 to 52.9%. From the four herds with high RR individuals, three farms dewormed the animals monthly, while the fourth farm dewormed the animals every 3 months. In farms where animals were dewormed every 6 months, low percentages of RR individuals were found, whereas no RR individuals were discovered in herds where animals were dewormed annually. These results suggest that the F200Y SNP is more significant in BZ resistance development of the surveyed population compared with the F167Y SNP.

Originally described in cattle, conglutinin belongs to the collectin family and is involved in innate immune defense. It is thought that conglutinin provides the first line of defense by maintaining a symbiotic relationship with the microbes in the rumen while inhibiting inflammatory reactions caused by antibodies leaking into the bloodstream. Due to the lack of information on the similar lectins and sequence detection in goats, we characterized the goat conglutinin gene using RACE and evaluated the differences in its gene expression profile, as well as in the gene expression profiles for surfactant protein A, galectins 14 and 11, interleukin 4 and interferon-gamma in goats. We used Saanen and Anglo Nubian F2 crossbred goats monitored over a period of four months and characterized them as resistant (R) or susceptible (S) based on the average values of EPG counts. Goat conglutinin was similar to bovine conglutinin, but its gene expression varied among different tissues. However, as with bovine conglutinin, it was most highly expressed in the liver. Variation in conglutinin (R = 24.3 ± 3.9; S = 23.5 ± 2.6, p = 0.059), protein surfactant A (R = 23.8 ± 5.2, S = 24.4 ± 2.3, p = 0.16), galectin 14 (R = 15.9 ± 3.5, S = 14.7 ± 6.2, p = 0.49) and galectin l1 gene expression (R = 25.4 ± 2.6, S = 25.8 ± 3.7, p = 0.53) was not significant between groups. However, there were weak correlations between interleukin 4 and the protein surfactant A gene (r = 0.459, p = 0.02) and between interleukin 4 and galectin 11 (r = 0.498, p = 0.01). Strong correlation between interferon-gamma and galectin 14 (r = 0.744, p = 0.00) was observed. Galectin 14 was negatively correlated with the number of nematodes in the goat (r = − 0.416, p = 0.04) as well as the EPG count (r = − 0.408, p = 0.04). This is the first study to date that identifies the gene expression of conglutinin, surfactant protein A and galectins 14 and 11 in the goat abomasum. In conclusion, we present evidence that lectin is involved in the immune response to gastrointestinal nematodes, which suggests that collectins and galectins are involved in the molecular recognition of helminths.

A polymerase chain reaction-based method was used to screen sandflies for infection with Wolbachia (Rickettsiales: Rickettsiaceae), an intracellular bacterial endosymbiont found in many arthropods and filarial hosts. Positive results were obtained in five of 200 field-collected sandflies and were confirmed by sequencing. All sandflies were Lutzomyia longipalpis (Diptera: Psychodidae) captured in a region endemic for visceral leishmaniasis in Brazil. This is the first study to identify Wolbachia infection in this Lutzomyia species, which is the main vector of leishmaniasis in the study area. The low infection rate found in this study (2.5%), together with the lack of detection of Wolbachia in previous studies and the diversity found in the sequences analysed, suggests horizontal transmission to these sandflies.

Benzimidazoles are the most common anthelminthic used for control of gastrointestinal nematodes of goats in the Brazilian semi-arid region. Resistance to these compounds in the nematode Haemonchus contortus has been associated with single nucleotide polymorphisms (SNP) in codons 167 (F167Y) and 200 (F200Y) on the β-tubulin isotype 1 gene. To determine the resistance profile to benzimidazoles of populations of H. contortus of goats of Brazilian semi-arid region, larvae of 29 populations of these nematodes were individually genotyped by real time PCR using a Taqman assay. The percentage of larvae homozygous (RR) for SNP F200Y was relatively low (18.9%), particularly when compared to SNP F167Y (32.7%), indicating that the latter has more relevance in this region. However, the associations between these two SNP demonstrate percentages of resistance ranging from 34.7% to 100% between populations, being the highest percentages for homozygous individuals resistant for the mutation 167 and susceptible to mutation 200 (RR-F167Y/F200Y-SS: 26.7%), followed by combination of heterozygous for both mutations (F167Y-SR/F200Y-SR: 22.8%). These results indicate high levels of resistance in populations of H. contortus of goats in the Brazilian semi-arid region, and thus ineffective antiparasitic control with the use of benzimidazoles in the region.

ContextCow milk is consumed in many countries. However, its consumption can be associated with gastrointestinal disorders in humans. The genetic profile of β-casein is one of the factors responsible for these disorders and the A1 variant is a causal agent. The β-casein A1 leads to the formation of bioactive peptides, which trigger intestinal reactions. The same is not observed for the A2 allele. A2 milk is obtained from cows with the A2A2 genotype. The identification of breeds and selection of animals for A2 milk production is important for public health and animal production. The Sindhi cattle (Bos taurus indicus) possesses characteristics such as adaptation to hot climate, resistance to parasites and utilisation of low-quality forage that are important for rearing in the tropics. AimsThe objective of the present study was to genotype the single-nucleotide polymorphism (SNP) that forms A1 and A2 alleles of the β-casein in Sindhi cattle. MethodsIn total, 348 Sindhi animals were used. DNA was extracted from hair follicles and submitted to PCR-sequencing for genotyping. Key resultsAllele frequency for A2 was 0.94 and for A1 it was 0.06. Genotype frequencies were 0.90 (A2A2), 0.09 (A1A2) and 0.01 (A1A1). The frequency of the A2A2 genotype in Sindhi cattle was similar to or higher than those found in zebu breeds and higher than those reported for specialised taurine breeds. ConclusionsThis high frequency of the A2A2 genotype indicates a genetic potential of the breed for A2 milk production and easy, assisted selection (if that is the intention). ImplicationsThis information gives importance to the breed and permits to add value to dairy product derived from it. It may increase producer’s income through producing and marketing A2 milk.

Cattleya elongata is a rupicolous orchid species spread throughout and endemic to outcrop islands in campo rupestre vegetation of the Chapada Diamantina, northeastern Brazil. We scored nine natural populations of C. elongata for morphological and genetic variability, covering the whole distribution area of the species, using allozymes and ISSR markers and morphometric multivariate analyses. Genetic variability in allozimes was relatively high (H e = 0.12–0.25), and unexpectedly higher than the values based on ISSR (H e = 0.16–0.19). The populations present moderate structuring (allozymes, ΦPT = 0.14; ISSR, ΦPT = 0.18) and low inbreeding (allozymes, F IS = 0.06). Genetic similarity among the populations was high in both markers, in spite of the discontinuity of the outcrops of the Chapada Diamantina. We found no particular biogeographical pattern to the distribution of the genetic and morphologic similarity among the populations of C. elongata. We found high morphological variability with moderate differentiation among the populations. We did not find any correlation among genetic, morphological, and geographical distances, and among the variability found in the morphological and genetic markers. The differences observed between the two genetic markers and the various morphological markers examined here indicated that the isolated use of any single parameter of these different populations for conservation planning or management would not consider all of the variability to be found in the species, as found in other Brazilian campos rupestres plants.

We carried out genetic and morphometric analyses in 33 populations belonging to all nine described and one putative taxa of Chamaecrista subsect. Baseophyllum. Genetic variability was low in all taxa. Morphological and genetic data are consistent with the status of C. depauperata and C. coriacea. However, divergence between C. cytisoides var. cytisoides and all the other six conspecific varieties is higher than the divergence between it and the other two species. Our data support the recognition of eight species in the subsection Baseophyllum. Chamaecrista coriacea and C. depauperata are supported in their original circumscription. However, the varieties ascribed to C. cytisoides are best treated as six different species. We propose five new combinations in order to raise varieties of C. cytisoides to specific rank: C. blanchetii, C. brachystachya, C. confertiformis, C. decora, C. unijuga. We also propose synonimization of one variety, C. cytisoides var. micrantha, under C. brachystachya.

Resumo A utilização de anti-helmínticos por longos períodos como principal medida de controle das parasitoses gastrintestinais de ruminantes levou a ineficácia aos levamisol, benzimidazóis e avermectinas. Este estudo descreve a atividade anti-helmíntica in vivo em populações naturais de nematoides trichostrongilídeos de caprinos. Foram selecionados 18 rebanhos provenientes dos biomas Caatinga (n=12) e Mata Atlântica (n=6), do Estado da Bahia, Brasil, criados em pastagens comunais em região semiárida. Grupos de oito a 10 animais foram tratados com albendazol (ABZ), ivermectina (IVM), levamisol (LEV), moxidectina (MOX) e closantel (CLOS). Os resultados do Teste de Redução da Contagem de Ovos nas Fezes indicaram resistência simultânea dos gêneros Haemonchus sp. e Trichostrongylus spp. para o ABZ, IVM, LEV, MOX e CLOS. As percentagens de eficácia variaram de 0-92%, 0-75%, 0-91%, 69-97% e 0-85% para o ABZ, IVM, LEV, MXD e CLOS, respectivamente, no bioma Caatinga e 0-59% para o ABZ e 9-59% para o IVM no bioma Mata Atlântica. Verificou-se nos rebanhos eficácia inferior a 95% para estes anti-helmínticos, com exceção de um único rebanho no qual a eficácia para MOX foi de 97%, o que sugere a presença de NGIs resistentes aos principais classes de anti-helmínticos em rebanhos caprinos destes biomas.

† Background and Aims Differences in the mating systems and the mechanisms of reproductive isolation between Chamaecrista desvauxii var. graminea and C. desvauxii var. latistipula were examined in the Chapada Diamantina, Brazil. These taxa occur sympatrically, and their populations demonstrate marked morphological differences. The objective of the present work was to determine if reproductive isolation mechanisms exist between these two populations of C. desvauxii, and to determine the influence of these putative mechanisms on their genetic differentiation. † Methods Field observations were made of floral biology, phenology and floral visitation, and experiments on intraand interpopulation pollination and germination rates of the resultant seeds were performed. A genetic examination of the populations was undertaken using four allozyme loci. † Key Results The varieties examined demonstrated overlapping of flowering periods during the months of June to September. The main pollinator for both varieties was the bee Bombus brevivillus. Both varieties are self-compatible, and a large number of fruits are formed in cross-pollinations with high seed germination rates. Inter-taxa pollinations result in high levels of fruit production, but no seeds are formed. Two of the four loci examined were diagnostic for the varieties, and exclusive high-frequency alleles were encountered at the other loci, leading to a high genetic distance between the two populations (0.495). † Conclusions Pre-zygotic barriers were not found between the two varieties, and these remain isolated due to postzygotic events. The two varieties demonstrate marked differences in their morphology, floral biology, phenology and genetic make-up, all of which indicate that they should be treated as two distinct species. A complete revision involving the other varieties of the C. desvauxii complex will be necessary in order to define these two taxa formally.

Melocactus glaucescens (Cactaceae) is a critically endangered species endemic to northeastern Brazil. It grows in sympatry with other congeneric species and there is evidence for hybridization among them. We evaluated the levels of genetic and morphological variability and their substructuring between populations of M. glaucescens and sympatric species, and we tested for the occurrence of natural hybridization. Genetic variability was investigated using 12 allozyme loci, and morphological variability was investigated using multivariate morphometric analyses of 18 vegetative characters in nine natural populations of M. glaucescens , Melocactus ernestii , Melocactus concinnus and two distinct morphs of putative hybrids ( Melocactus ¥ albicephalus ). Genetic variability was low in all populations ( P = 7.7–41.7, A = 0.3–1.7, H e = 0.009–0.096), and all taxa displayed a deficit in heterozygotes. Low genetic structuring and moderate morphological structuring were found for M. glaucescens ( F ST = 0.045, A MRPP = 0.16) and M. concinnus ( F ST = 0.022, A MRPP = 0.11). The results obtained are inconclusive with respect to confirming the hypothesis of occurrence of hybridization because of an absence of diagnostic loci in the presumed parental species. The presence of several private alleles and the absence of linkage disequilibrium in the putative hybrids indicate that M. albicephalus should be treated as a distinct species, and not given hybrid status as recently stated by a number of authors. The large number of exclusive alleles and the levels of morphological structuring in the populations of M. glaucescens are important factors that need to be considered in the definition of strategies for the conservation of this species.

Foi estudada a biologia reprodutiva de Melocactus glaucescens e M. paucispinus (Cactaceae) no Município de Morro do Chapéu, Chapada Diamantina, Bahia, sendo abordados aspectos da fenologia, biologia floral, polinização e sistema reprodutivo. Foram registrados os períodos de floração e frutificação, visitantes florais, freqüência e tipo de visitas, além de estratégia e comportamento dos visitantes às flores. Foram realizadas polinizações experimentais para verificar o sistema reprodutivo das espécies. As duas espécies de Melocactus estudadas apresentaram sobreposição de floração ao longo do período de estudo. Os atributos florais de ambas as espécies são típicos da síndrome da ornitofilia: cores atrativas, estrutura tubulosa e produção de néctar com baixa concentração de solutos, entre 20% e 30%. O beija-flor Chlorostilbon aureoventris Boucier & Mulsant (1948) foi o visitante mais freqüente, com 82% e 89% do total de visitas para M. paucispinus e M. glaucescens, respectivamente. Outras espécies de beija-flores e borboletas também visitaram as flores das espécies. A sobreposição no período de floração e a similaridade na composição da guilda de polinizadores destas e demais espécies simpátricas de Melocactus favorece a hibridação no gênero, como foi observado na região. Melocactus glaucescens apresentou auto-incompatibilidade e alogamia, enquanto M. paucispinus é auto-compatível e autogâmica, porém com menor frutificação em autopolinização do que em polinização cruzada, possivelmente devido à ocorrência de depressão endogâmica. Polinizações interespecíficas indicam a ocorrência de inter-compatibilidade entre M. paucispinus e M. concinnus Buining & Brederoo, sustentando hipóteses correntes de hibridação entre estas espécies.

• Background and Aims Melocacatus paucispinus (Cactaceae) is endemic to the state of Bahia, Brazil, and due to its rarity and desirability to collectors it has been considered threatened with extinction. This species is usually sympatric and inter-fertile with M. concinnus, and morphological evidence for hybridization between them is present in some populations. Levels of genetic and morphological variation and sub-structuring in populations of these species were assessed and an attempt was made to verify the occurrence of natural hybridization between them. • Methods Genetic variability was surveyed using allozymes (12 loci) and morphological variability using multivariate morphometric analyses (17 vegetative characters) in ten populations of M. paucispinus and three of M. concinnus occurring in the Chapada Diamantina, Bahia. • Key Results Genetic variability was low in both species (P = 0·0–33·3, A = 1·0–1·6, He = 0·000–0·123 in M. paucispinus; P = 0·0–25·0, A = 1·0–1·4, He = 0·000–0·104 in M. concinnus). Deficit of heterozygotes within the populations was detected in both species, with high values of FIS (0·732 and 0·901 in M. paucispinus and M. concinnus, respectively). Evidence of hybridization was detected by the relative allele frequency in the two diaphorase loci. High levels of genetic (FST = 0·504 in M. paucispinus and 0·349 in M. concinnus) and morphological (A = 0·20 in M. paucispinus and 0·17 in M. concinnus) structuring among populations were found. • Conclusions The Melocactus spp. displayed levels of genetic variability lower than the values reported for other cactus species. The evidence indicates the occurrence of introgression in both species at two sites. The high FST values cannot be explained by geographical substructuring, but are consistent with hybridization. Conversely, morphological differentiation in M. paucispinus, but not in M. concinnus, is probably due to isolation by distance.

Galectins and collectins are proteins classified in the lectin family that have the ability to recognize molecular patterns associated with pathogens. Studies on cattle have demonstrated high expression of these proteins during infection with gastrointestinal nematodes. The aim of this study was to investigate whether the level of Haemonchus contortus infection would alter the expression of galectins (Gal11 and Gal14) and collectins (SPA and CGN) in sheep. Twelve Corriedale sheep exposed to natural infection with nematodes were divided into two groups: group 1 (G1, n = 7) and group 2 (G2, n = 5), with low and high parasite burdens, respectively, based on fecal egg counts and abomasal parasite counts. The fecal egg counts and abomasal parasite counts were significantly different (p < 0.05) between the groups. Galectin and collectin gene expression was observed in all sheep abomasal samples. However, animals with lower infection levels showed lower expression of the genes Gal14, SPA and CGN (p < 0.05). Expression of lectins was associated with the abomasal H. contortus burden, thus suggesting that these proteins may have a role in controlling of this infection. Colectinas e galectinas são proteínas da família das lectinas que possuem a capacidade de reconhecer padrões moleculares associados aos patógenos. Estudos em bovinos têm demonstrado a alta expressão dessas proteínas durante a infecção por nematoides gastrintestinais. O objetivo deste estudo foi investigar se o nível de infecção de Haemonchus contortus altera a expressão de colectinas (SPA e CGN) e galectinas (Gal11 e Gal14) de ovinos. Doze ovinos da raça Corriedale expostos a infecção natural com nematoides foram separados em dois grupos: grupo 1 (G1, n=7) com menor grau de parasitismo; e grupo 2 (G2, n=5) com maior grau, a partir da contagem do número de parasitos recuperados do abomaso e OPG. A contagem de OPG e de parasitos recuperados do abomaso dos grupos G1 e G2 apresentaram diferença estatística (p