The order Chiroptera is considered the second largest group of mammals in the world, hosting important zoonotic virus and bacteria. Bartonella and hemotropic mycoplasmas are bacteria that parasite different mammals' species, including humans, causing different clinical manifestations. The present work aimed investigating the occurrence and assessing the phylogenetic positioning of Bartonella spp. and Mycoplasma spp. in neotropical bats sampled from Brazil. Between December 2015 and April 2016, 325 blood and/or tissues samples were collected from 162 bats comprising 19 different species sampled in five states of Brazil. Out of 322 bat samples collected, while 17 (5·28%) were positive to quantitative PCR for Bartonella spp. based on nuoG gene, 45 samples (13·97%) were positive to cPCR assays for hemoplasmas based on 16S rRNA gene. While seven sequences were obtained for Bartonella (nuoG) (n = 3), gltA (n = 2), rpoB (n = 1), ftsZ (n = 1), five 16S rRNA sequences were obtained for hemoplasmas. In the phylogenetic analysis, the Bartonella sequences clustered with Bartonella genotypes detected in bats sampled in Latin America countries. All five hemoplasmas sequences clustered together as a monophyletic group by Maximum Likelihood and Bayesian Inference analyses. The present work showed the first evidence of circulation of Bartonella spp. and hemoplasmas among bats in Brazil.
This study examined the development of resistance to anthelmintics in Haemonchus contortus in lambs under suppressive or selective treatment regimens that included monepantel. Twenty Ile de France and 20 Santa Ines lambs were allocated to two anthelmintic treatment regimens, based on body weight and nematode faecal egg counts (FEC): targeted selective treatment (TST) or suppressive treatment, both with monepantel. Lambs of the TST group were treated individually when they presented with a packed cell volume (PCV) ≤20%. On 7 October 2016, the lambs were allocated to clean pastures, where they grazed in separated paddocks by group until late February 2017. The experimental area was contaminated with nematodes that were introduced with the experimental Ile de France and Santa Ines lambs, naturally infected with gastrointestinal nematodes. To maintain the grazing lambs in the suppressive treatment group and their pasture as free of worms as possible, these lambs were treated with anthelmintics before being allocated to their paddock and then were periodically treated with monepantel. However, the use of a suppressive treatment regimen that included monepantel over a period of 3 months resulted in the emergence of a population of resistant H. contortus. In the TST group, there was a rapid and progressive reduction in the efficacy of monepantel, which at the end of the experiment was only 76%. The Ile de France lambs were all treated one or more times during the experiment, whereas only two Santa Ines lambs in the TST required treatment. In conclusion, a population of H. contortus resistant to monepantel emerged quickly during the rainy season, even when sheep were submitted to selective treatment.
Understanding the immunological basis of resistance to gastrointestinal nematode infections in livestock is important in order to develop novel methods of parasite control such as vaccination or genetic selection for parasite resistance. The present study aimed to investigate differences in immune response between parasite resistant Santa Ines and susceptible Ile de France sheep breeds to natural Haemonchus contortus infection. Parasitological parameters, humoral immunity, local and circulating cellular immune responses were evaluated in 19 Santa Ines and 19 Ile de France lambs undergoing different anthelmintic treatments regimens: suppressive treatments (SUP) or targeted selective treatments (TST) over a 5-month grazing period. Santa Ines lambs had significantly lower Haemonchus faecal egg count and worm burden compared to Ile de France regardless of treatment regime. In addition, circulating blood eosinophils count and parasite-specific IgG levels were significantly higher and more rapidly induced in Santa Ines lambs. Abomasal immune responses were generally greater in the resistant breed, which had significantly higher levels of parasite-specific IgA in mucus, and elevated number of globule leukocytes and CD3+ T cells within the abomasal mucosal. Furthermore, numbers of POU2F3+ epithelial cells, a tuft-cell specific transcription factor, were also elevated in the Santa Ines breed, suggesting that this breed is better able to initiate T-helper type 2 immune responses within the abomasum. In conclusion, the differential immunological responses detailed here are relevant to understanding resistance to gastrointestinal nematodes in other host breeds, as well as to resistance breeding as a sustainable control approach for parasitic infections.
Lamb performance was evaluated in four integrated crop-livestock (ICL) systems. The ICL areas were previously planted with maize and marandu palisade grass in December 2013. ICL systems 1 and 2 were also seeded with pigeon pea. After harvesting in April 2014, black oats were sown in rows (ICL 1 and 3) or by broadcast seeding (ICL 2 and 4). Each ICL area was divided into 12 paddocks of 225 m 2 to be grazed by young sheep from July 23 to September 30, 2014. To determine if the pasture remained contaminated by free-living stages of sheep gastrointestinal nematodes (GIN) after approximately ten months without animals in the area, 12 worm-free "tracer" lambs (n = 3/ ICL system) grazed each ICL pasture for 14 consecutive days in July 2015 and were later housed in pens, where they remained for another 14 days. The tracer lambs acquired no worm infections, demonstrating that the area was cleared after 300 days without animals. To evaluate GIN infection and uncastrated male lamb performance, we used 60 (n = 15/ system) Poll Dorset x Corriedale (crossbred) sheep with mean body weights of 24.4 ± 3.4 kg from July to September 2015 and 48 (n = 12/ system) Texel x Corriedale sheep with body weights of 26.4 ± 3.5 kg from June to September 2016. Lambs were allocated to the following groups: Group 1 rotated on the 12 ICL1 paddocks; Group 2 rotated on the 12 ICL2 paddocks; Group 3 rotated on the 12 ICL3 paddocks; and Group 4 rotated on the 12 ICL4 paddocks. Each paddock was grazed by sheep twice for three days, with a 33-day interval between grazing. At the end of the day the lambs were supplemented with concentrate plus silage. Groups 1 and 2 received mixed silage made of maize, marandu palisade grass and pigeon pea and groups 3 and 4 received mixed silage made of maize and marandu palisade grass. Faecal and blood samples were taken from all the animals every two weeks, and body weight was recorded on the same occasion. Nematode faecal egg counts (FEC), packed cell volume and total plasma protein means did not differ (P > 0.05) between the four lamb groups. In 2015, throughout the experimental period, the FEC decreased in all animals, with 6733 and 1407 eggs per gram (EPG) on average at the beginning and end of the trial, respectively. At the beginning of the trial in 2016, the animals had mild GIN infections (1077 EPG on average), and the individual faecal egg counts did not exceed 10,000 EPG during the trial. In decreasing order, Haemonchus spp., Trichostrongylus spp. and Cooperia spp. third stage larvae were found in the faecal cultures in both years. The performance of lambs were similar in the four ICL systems (P > 0.05), in the first and second years, the daily weight gains were 0.192 ± 0.05 kg and 0.221 ± 0.06 kg, respectively. In conclusion, a period of withdrawal of contaminated sheep results in plots free of infective larvae. The use of clean pastures during the dry season, in the different ICL systems, associated with a good nutrition plan, resulted in progressively declining degrees of GIN infections and satisfactory ...
Background Haemonchus contortus , a gastrointestinal nematode parasite of sheep, is mainly controlled by anthelmintics; the occurrence of anthelmintic resistance leads to treatment failures and increases economic burden. Because molecular mechanisms involved in drug resistance can be elucidated by genomic studies, an extreme quantitative trait locus (X-QTL) mapping approach was used to identify co-segregation of the resistance phenotype with genetic markers to detect the genome-wide variants associated with monepantel resistance in H. contortus . Methods A cross between H. contortus isolates using parental susceptible (Par-S) males and monepantel resistant (Par-R) females resulted in SR progeny, while reciprocal cross resulted in RS progeny. Pools ( n = 30,000) of infective larvae (L3) recovered from Par-R, and from SR and RS populations in the F3 generation, collected both before (unselected group) and 7 days after (selected group) selection with monepantel treatment in sheep hosts, were subjected to genome sequencing (Pool-Seq). Pairwise comparisons of allele frequencies between unselected and selected groups were performed for each population by Fisher’s exact test (FET) and for both populations combined by a Cochran-Mantel-Haenszel (CMH) test. Results Mapping rates varied from 80.29 to 81.77% at a 90.4X mean coverage of aligned reads. After correction for multiple testing, significant ( P < 0.05) changes in allele frequencies were detected by FET for 6 and 57 single nucleotide polymorphisms (SNPs) in the SR and RS populations, respectively, and by the CMH test for 124 SNPs in both populations. The significant variants located on chromosome 2 generated a selection signal in a genomic region harboring the mptl-1, deg-3 and des-2 genes, previously reported as candidates for monepantel resistance. In addition, three new variants were identified in the mptl-1 gene. Conclusions This study expands knowledge on genome-wide molecular events underlying H. contortus resistance to monepantel. The identification of a genome region harboring major genes previously associated with monepantel resistance supports the results of the employed X-QTL approach. In addition, a deletion in exon 11 of the mptl-1 gene should be further investigated as the putative causal mutation leading to monepantel resistance. Electronic supplementary material The online version of this article (10.1186/s13071-019-3663-9) contains supplementary material, which is available to authorized users.
Amazonia, the largest Brazilian biome, is one of the most diverse biomes around the world. Considering the Brazilian chiropteran species, 120 out of known 167 species are registered in Pará state, with 10 endemic species. Despite the high diversity of bats in Amazonia, studies on their parasites, especially on helminths, are scarce. Therefore, the present study aims to study the helminth fauna of different bats from the Pará state, Amazon biome, determine the descriptors of infection, and evaluate the host-parasite interactions, as well as evaluate differences in ecological indexes in accordance with the feeding guilds. The study was developed on 67 bats of 21 species captured in several areas of the Pará state. The animals were identified, divided into feeding guilds, and necropsied. The parasites obtained were identified and quantified. A total of 182 parasites were found in 20.89 % of the studied bats, representing nine species, as follows: Anenterotrema eduardocaballeroi, Anenterotrema liliputianum, Ochoterenatrema caballeroi, Tricholeiperia sp., Parahistiostrongylus octacanthus, Litomosoides guiterasi, Litomosoides brasiliensis, Capillariinae gen. sp., and Hymenolepididae gen. sp. Also, the results indicated that there was no impact of parasitism on host body condition and no relationship between sex and parasite intensity. In relation to the feeding guilds, the omnivores showed higher prevalence and mean intensity. Animals from regions closer to the equator tend to have greater richness in parasite species, but the present study revealed low diversity and richness in species. In conclusion, the ecological pattern observed for other animal groups, in which higher parasitic diversity are registered in lower latitudes, is not applicable to chiropterans from the study area.
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