Two in vitro anthelmintic assays of four Paraguayan medicinal plants for proof of concept of the role of polyphenols in their biological activities and LC-HRMS analysis.

ETHNOPHARMACOLOGICAL RELEVANCE: Gastrointestinal nematodes (GIN) control in small ruminants has relied on the systematic use of synthetic anthelmintics (AH), their effectiveness has been progressively decreasing due to the rise and diffusion of anthelmintic resistances. The most prevalent genera affecting small ruminants were Haemonchus spp., and Trichostrongylus spp. The investigation of new anthelmintics in plants is a highly studied option, especially when it is linked to ethnobotanical knowledge and phenolic compounds. THE AIMS OF OUR STUDY: Four medicinal plants mentioned in traditional medicine were selected to evaluate their anthelmintic proprieties at different stages of the life cycle of GIN, namely: Kyllinga odorata Valh., Cassia occidentalis L., Artemisia absinthium L, and Verbena litoralis Kunth and to explore the role of polyphenols in the AH activity. MATERIALS AND METHODS: To evaluate the anthelmintic activity in this study, two models of GIN species, namely Haemonchus contortus (Hc) and Trichostrongylus colubriformis (Tc) were selected and tested on two in vitro assays: 1) Larval Exsheathment Inhibition Assay (LEIA) and, 2) Egg Hatch Assay (EHA). To explore the role of tannins and polyphenols in AH activity by comparing the effects of LEIA and EHA with or without polyvinylpolypyrrolidone (PVPP) and to characterize the phytochemical composition of the most active plants using ultra-high performance liquid chromatography (UHPLC) coupled with high-resolution mass spectrometry (HRMS). RESULTS: C. occidentalis exhibited the highest activity on LEIA (EC50 = 250.42-41.80 µg/mL) and A. absinthium on egg hatching processes (EC50 = 121.70-137.34 µg/mL) in both species of GIN. The inhibition in the development of eggs was from 67.70% to 96.36% on H. contortus, and from 78.87% to 99.65% on T. colubriformis. At the maximal dose, Additionally, it was observed that the AH on eggs varies according to the GIN species: on H. contortus the extracts tested blocked the formation of larvae Ovicidal Effect (% higher OE) and on T. colubriformis they blocked the appearance of L1 larvae, Larvae Failing Eclosion (% higher LFE). After PVPP, a reduction in AH activity on LEIA and EHA was noted, especially with C. occidentalis (87.20-67.00% of larvae exsheathment, (p < 0.05) and 40.51-24.96% of egg hatching, (p > 0.05) of both parasite species. Nine putative features were identified using HRMS and MS/MS after addition of PVPP. CONCLUSIONS: The present study demonstrated that C. occidentalis, A. absinthium, and K. odorata, which parts have been traditionally used as medicinal plants are a valuable source of active compounds with anthelmintic activity. The medicinal use of these plants against GIN parasites was proven by in vitro analysis. Therefore exploration of the secondary metabolites of these plant extracts and testing of isolated fractions of active compounds under in vivo experiments are planned and represent a specific challenge for alternative drug research. Regarding the PVPP, in this study we hypotheses about the standard doses it was not able to completely absorb the polyphenols of extracts of K. odorata, C. occidentalis, and A. absinthium, which would lead to more studies to evaluate the role of this product in the absorption of phenolic compounds.

Influence of targeted selective anthelmintic treatment on the productive performance of wool and hair lambs naturally infected with gastrointestinal nematodes in Brazil.

Targeted selective treatment (TST) is an alternative method to reduce the use of anthelmintics and delay the development of resistant nematode populations. However, there is limited information on the actual effects of this type of treatment on livestock productivity. The objective of this study was to evaluate the production performance of Santa Ines (hair) and Ile de France (wool) lambs naturally infected with gastrointestinal nematodes (GIN) under TST based on packed cell volume (PCV) versus suppressive anthelmintic treatments. Thirty-eight lambs were divided into two treatment groups: Suppressive treatment, animals were drenched with monepantel every two weeks and TST, animals were treated with the same anthelmintic when they presented PCV ≤ 20%. Feces, blood, and weight were measured weekly to determine eggs per gram of feces, PCV, total plasma protein, and weight gain. After animals were slaughtered, carcasses were weighed to determine carcass yield. In the TST group, substantial productive losses of approximately 21.3% in the wool and 25.9% in the hair lambs were observed in body weight compared to their counterparts. Significant differences in hematological variables occurred over the experimental period, especially in the wool lambs under TST. Favorable environmental conditions enabled infective larvae to survive and thrive on pasture. Haemonchus contortus and intestinal nematodes were the most common parasites found in the Ile de France lambs and the Santa Ines lambs, respectively. Although TST prevented mortality, it did not prevent production losses. Both breeds showed a significant drop in production due to GIN parasitism.

Gastrointestinal Parasites in Sheep from the Brazilian Pampa Biome: Prevalence and Associated Factors.

Parasitic infections are common in sheep farming worldwide. A cross-sectional study was carried out to determine the prevalence and associated risk factors among gastrointestinal parasites and sheep herds from the Brazilian Pampa biome, Rio Grande do Sul state, Brazil. Twenty-one farms were visited, where rectal faecal samples were individually collected from 309 animals. The sheep industry on the studied farms was characterised by small farms with a low level of technification, an extensive grazing system and meat production. Of all samples analysed, strongyle nematodes had the highest prevalence (77.02%), followed by Eimeria spp. (70.55%), Moniezia expansa (20.39%) and Strongyloides papillosus (17.48%). Mixed infection, between helminths and protozoans, was detected in 68.61% of samples. All herds were positive for strongyle and Eimeria spp. A predominance of Haemonchus spp. and Trichostrongylus spp. nematodes was observed in the herds. Younger animals were significantly more affected by Eimeria and M. expansa. In the semi-intensive and intensive systems, a higher frequency of Eimeria and strongyle infections was observed. Parasite infection was significantly reduced at low animal densities. The Brazilian Pampa region presents a high prevalence of gastrointestinal parasites among sheep; age, breeding system and stocking density were factors associated with parasite infection.

As infecções parasitárias são comuns na criação de ovinos em todo o mundo. Um estudo transversal foi realizado para determinar a prevalência e fatores de risco associados entre parasitoses gastrintestinais e rebanhos ovinos do bioma Pampa brasileiro, estado do Rio Grande do Sul, Brasil. Vinte e uma fazendas foram visitadas, onde foram coletadas amostras de fezes individuais da ampola retal de 309 animais. A produção ovina era caracterizada por pequenas propriedades com baixo nível de tecnificação, os animais eram criados em sistema extensivo e para produção de carne. De todas as amostras analisadas, os estrongilídeos (Nematoda: Strongylida) foram mais prevalentes (77,02%), seguido por Eimeria spp. (70,55%), Moniezia expansa (20,39%) e Strongyloides papillosus (17,48%). Infecção mista, entre helmintos e protozoários, foi detectada em 68,61% das amostras. Todos os rebanhos foram positivos para estrongilídeos e Eimeria spp. Foi observada predominância dos nematódeos Haemonchus spp. e Trichostrongylus nos rebanhos. Animais mais jovens foram significativamente mais afetados por Eimeria e M. expansa. Nos sistemas semi-intensivo e intensivo, observou-se maior frequência de infecções por Eimeria e estrongilídeos. A infecção parasitária foi significativamente menor em fazendas com baixa densidade animal. A região do Pampa brasileiro apresenta alta prevalência de parasitos gastrintestinais em ovinos; idade, sistema reprodutivo e densidade animal foram fatores associados à infecção parasitária.

Mortality caused by gastrointestinal nematodes in beef cattle submitted to an inadequate sanitary protocol / Mortalidade causada por nematódeos gastrintestinais em bovinos de corte submetidos a protocolo sanitário inadequado

The aim of this study was to describe the epidemiological, clinical, and pathological findings of two outbreaks of gastrointestinal nematode infections in beef cattle kept under an inadequate sanitary protocol. An outbreak of trichostrongylosis occurred in Amambai, Mato Grosso do Sul (MS), from May to July 2016. The herd consisted of 3,000 Nellore cows, and of these, fifteen died after showing emaciation and diarrhea and remaining in recumbency. At necropsy, the abomasum showed edema in the folds, white, raised areas, multifocal to coalescent, and small ulcers. Histopathology showed larvae compatible with trichostrongylid nematodes were present inside the abomasum glands. Trichostrongylus axei was identify in the abomasum. The hemonchosis outbreak began in October 2018, in a breeding and finishing property in Santa Rita do Pardo, MS. Of 4,000 cattle aged 8 to 18 months, 673 became ill and 117 died. Clinical signs were emaciation, weakness, dehydration, submandibular edema, and soft stools. At necropsy, large numbers of nematodes were found in the abomasum; they were morphologically classified as Haemonchus placei. Both outbreaks were caused by failures in the parasite control protocol. However, in Outbreak I, frost and immune stress caused by lack of food may have contributed to the deaths. In Outbreak I, the main failure in the devermination protocol was the use of anthelmintics without carrying out an efficacy test. In Outbreak II, there was no parasite control protocol in the rearing and finishing property, and in the breeding property, there was resistance to the anthelmintic used (ivermectin). The two outbreaks highlight the importance of gastrointestinal nematode control protocols in cattle and demonstrate that infections by T. axei and H. placei can be lethal for this species.

O objetivo deste trabalho foi descrever os achados epidemiológicos, clínicos e patológicos de dois surtos de verminose em bovinos de corte mantidos sob protocolo sanitário inadequado. Um surto de tricostrongilose ocorreu em Amambai, Mato Grosso do Sul (MS), de maio a julho de 2016. O rebanho de cria era composto por 3.000 vacas Nelore, das quais, quinze vacas adultas morreram após apresentarem emagrecimento, diarreia e permanecerem em decúbito. Na necropsia, o abomaso apresentava edema das pregas, áreas brancacentas, elevadas, multifocais a coalescentes e pequenas úlceras. Na histopatologia, larvas compatíveis com nematódeos tricostrongilídeos estavam presentes no interior das glândulas do abomaso. A recuperação de helmintos do abomaso demonstrou que Trichostrongylus axei foi o principal agente etiológico. O surto de hemoncose iniciou-se em outubro de 2018, em uma propriedade de recria e terminação em Santa Rita do Pardo, MS. Do total de 4 mil bovinos com 8 a 18 meses, 673 adoeceram e 117 morreram. Os sinais clínicos foram emagrecimento, fraqueza, desidratação, edema submandibular e fezes pastosas. Na necropsia, havia grande quantidade de nematódeos no abomaso, que foram classificados morfologicamente como Haemonchus placei. Ambos os surtos foram causados por falhas no protocolo de controle parasitário, porém, no Surto I, geadas e estresse imunológico causado pela falta de alimento podem ter contribuído para as mortes. No Surto I, a principal falha no protocolo de desverminação foi o uso de anti-helmínticos sem a realização de teste de eficácia. No Surto II, não havia protocolo de controle parasitário na propriedade de recria e terminação, e na propriedade de cria, havia resistência ao anti-helmíntico utilizado (ivermectina). Os dois surtos ressaltam a importância dos protocolos de controle de nematódeos gastrintestinais em bovinos e demonstram que infecções por T. axei e H. placei podem ser letais para esta espécie.

Brote familiar por el nematodo Trichostrongylus colubriformis en una zona rural de la provincia de Valdivia: una zoonosis de rara ocurrencia / Family outbreak caused by the nematode Trichostrongylus colubriformis in a rural area of the province of Valdivia: a rare occurrence zoonoses

Resumen La trichostrongylosis es una zoonosis de rara ocurrencia causada por nematodos intestinales, favorecida por el consumo de vegetales crudos o agua contaminada con larvas infectantes de Trichostrongylus spp. En 2015, se registró un brote familiar transmitido por alimentos en una zona rural de Valdivia, afectando a una mujer de 51 años de edad y su pareja e hija de 56 y 12 años; respectivamente. Solo la mujer adulta, el caso índice, presentó síntomas como diarrea, dolor abdominal, astenia y náuseas. Los pacientes eliminaron huevos de Trichostrongylidae gen. sp. en sus deposiciones, identificándose en dos de ellos, adultos de Trichostrongylus colubriformis. Tuvieron una buena respuesta a albendazol. En Chile, se han registrado prevalencias de infección de 0,1 a 3,5%, incluyendo un total de 93 casos diagnosticados entre las provincias de Concepción y Llanquihue.

Abstract Trichostrongylosis is a rare occurrence zoonosis caused by intestinal nematodes, favored by the consumption of raw vegetables or water contaminated with infective larvae of Trichostrongylus spp. In 2015, a family outbreak of foodborne disease was registered in a rural zone of Valdivia, affecting to 51-year-old woman and her 56-year-old partner and her 12-year-old daughter. Only the adult woman, the index case, presented symptoms such as diarrhea, abdominal pain, asthenia, and nausea. The patients revealed Trichostrongylidae gen. sp. eggs in their stools, identifying Trichostrongylus colubriformis adults in two of them. They had a good response to treatment with albendazol. In Chile, 0.1 to 3.5% prevalence of infection has been reported, including a total of 93 cases recorded between the provinces of Concepción and Llanquihue.

Isolation of pure Trichostrongylus colubriformis strains from naturally infected sheep using two methodologies.

Two methodologies were tested to isolate pure Trichostrongylus colubriformis strains from naturally infected sheep. Also, the in vitro susceptibility status to commercial anthelmintic (AH) drugs was determined in these strains. These methods were performed in a tropical region of Mexico where naturally infected sheep and goats host Haemonchus contortus, T. colubriformis and Oesophagostomum columbianum. For the first strain, a group of 6 naturally infected lambs from the "Paraiso" farm were treated with closantel (subcutaneous (SC), 10 mg/kg). On day 10 post-treatment, the eggs per gram (EPG) of faeces were determined with the McMaster technique. The faeces from the two lambs with the highest EPG were used for coprocultures (4 days, 28 °C). The L3 larvae were recovered and identified as T. colubriformis (69%) and O. columbianum (31%). The latter was removed by 10-day refrigeration (4-5 °C) followed by sieving (25 µm), resulting in 100% T. colubriformis (PARAISO strain). The second strain was isolated using repeated doses of levamisole (LEV, SC 7.5 mg/kg) in an 8-year-old sheep. The sheep had 1700 EPG before the LEV treatments and maintained 1300 EPG after both LEV treatments (day 10). The coproculture (4 days, 28 °C) after the second treatment contained 100% T. colubriformis (FMVZ-UADY strain). The in vitro AH susceptibility was determined using the egg hatch test for benzimidazole (BZ), and the larval migration inhibition test for ivermectin (IVM) and LEV. The PARAISO strain was BZ- and LEV-susceptible, and IVM-resistant. Meanwhile, the FMVZ-UADY strain was BZ- and IVM-susceptible, and LEV-resistant. The conditions where these two protocols could be used in other parts of the world were discussed.

First Molecular Characterization of Trichostrongylus colubriformis Infection in Rural Patients from Chile.

PURPOSE: The aim was to characterize the infection by Trichostrongylus spp. in patients from Chile using a combination of molecular detection techniques and phylogenetic analysis relating the findings to clinical and epidemiological reports of the patients METHODS: Strongylid eggs were detected in seven patients by coproparasitological techniques. From each sample a fragment of the ITS-2 ribosomal gene was amplified by PCR, sequenced and analyzed by the Neighbor-Joining method. RESULTS: All the sequences and phylogenetic clusters corresponded to T. colubriformis. Two samples presented a single nucleotide polymorphism showing two possible haplotypes. Six patients presented gastrointestinal symptoms. All of them lived on farms and used sheep manure as fertilizer. CONCLUSION: T. colubriformis was the strongylid involved in the infections of these Chilean patients associated with the presence of livestock and agricultural practices that favor infection by this type of nematode.

Losses caused by gastrointestinal nematode infections in Dorper lambs under two nutritional status / Prejuízos causados pelas infecções por nematoides gastrintestinais em cordeiros Dorper submetidos a dois manejos nutricionais

Abstract The aim of this study was to evaluate the effect of two nutritional statuses on the productive performance of Dorper lambs naturally infected with gastrointestinal nematodes. Thirty-two lambs, grazing together on the same pasture, were allocated into four experimental groups: (G1) infected-supplemented diet, (G2) control-supplemented diet, (G3) infected-basal diet, and (G4) control-basal diet. Control animals received suppressive treatment with monepantel every two weeks, while precautionary anthelmintic treatments were given to all lambs of the infected groups with packed cell volume (PCV) <23%. There was reduction in the PCV means of all groups, which was more pronounced in the infected lambs that also presented reduction in total plasma protein values in comparison with the controls. Weight gain was affected by diet and infection status (P < 0.05). Daily body weight gain was 0.170 kg in the G1, 0.205 kg in the G2, 0.085 kg in the G3, and 0.116 kg in the G4. The cold carcass weight was 4.1% and 13.7% higher in controls in comparison with infected lambs, respectively, in the supplemented and basal diets. The infected groups, despite receiving precautionary anthelmintic treatments to prevent deaths due to haemonchosis, presented reduction in the production parameters in comparison with the controls.

Resumo O experimento teve por objetivo determinar o efeito de dois níveis de nutrição no desempenho produtivo de cordeiros Dorper naturalmente infectados por nematoides gastrintestinais. Trinta e dois cordeiros, mantidos juntos na mesma pastagem, foram alocados em quatro grupos experimentais: (G1) infectado-suplementado, (G2) controle-suplementado, (G3) infectado-dieta basal e (G4) controle-dieta basal. Os cordeiros suplementados receberam diariamente concentrado em quantidade equivalente a 2% do peso corporal (PC), enquanto na dieta basal receberam apenas uma pequena quantidade de concentrado (0,35% do PC). Os animais controles receberam tratamento supressivo com anti-helmíntico a cada duas semanas e os infectados foram tratados individualmente quando apresentaram volume globular (VG) <23%. Houve redução nas médias de VG em todos os grupos, as quais foram mais pronunciadas nos animais dos grupos infectados, que também apresentaram redução nos valores de proteína plasmática total em comparação com os controles. Houve efeito significativo da dieta e da infecção no ganho de peso (P <0,05). O ganho em peso diário foi de 0,170 kg no G1, 0,205 kg no G2, 0,085 kg no G3 e 0,116 kg no G4. Os grupos infectados, apesar de receberem tratamentos anti-helmínticos preventivos que evitaram mortes por haemonchose, apresentaram redução nos parâmetros produtivos em comparação com os controles.

Losses caused by gastrointestinal nematode infections in Dorper lambs under two nutritional status / Prejuízos causados pelas infecções por nematoides gastrintestinais em cordeiros Dorper submetidos a dois manejos nutricionais

The aim of this study was to evaluate the effect of two nutritional statuses on the productive performance of Dorper lambs naturally infected with gastrointestinal nematodes. Thirty-two lambs, grazing together on the same pasture, were allocated into four experimental groups: (G1) infected-supplemented diet, (G2) control-supplemented diet, (G3) infected-basal diet, and (G4) control-basal diet. Control animals received suppressive treatment with monepantel every two weeks, while precautionary anthelmintic treatments were given to all lambs of the infected groups with packed cell volume (PCV) 23%. There was reduction in the PCV means of all groups, which was more pronounced in the infected lambs that also presented reduction in total plasma protein values in comparison with the controls. Weight gain was affected by diet and infection status (P 0.05). Daily body weight gain was 0.170 kg in the G1, 0.205 kg in the G2, 0.085 kg in the G3, and 0.116 kg in the G4. The cold carcass weight was 4.1% and 13.7% higher in controls in comparison with infected lambs, respectively, in the supplemented and basal diets. The infected groups, despite receiving precautionary anthelmintic treatments to prevent deaths due to haemonchosis, presented reduction in the production parameters in comparison with the controls.(AU)

O experimento teve por objetivo determinar o efeito de dois níveis de nutrição no desempenho produtivo de cordeiros Dorper naturalmente infectados por nematoides gastrintestinais. Trinta e dois cordeiros, mantidos juntos na mesma pastagem, foram alocados em quatro grupos experimentais: (G1) infectado-suplementado, (G2) controle-suplementado, (G3) infectado-dieta basal e (G4) controle-dieta basal. Os cordeiros suplementados receberam diariamente concentrado em quantidade equivalente a 2% do peso corporal (PC), enquanto na dieta basal receberam apenas uma pequena quantidade de concentrado (0,35% do PC). Os animais controles receberam tratamento supressivo com anti-helmíntico a cada duas semanas e os infectados foram tratados individualmente quando apresentaram volume globular (VG) 23%. Houve redução nas médias de VG em todos os grupos, as quais foram mais pronunciadas nos animais dos grupos infectados, que também apresentaram redução nos valores de proteína plasmática total em comparação com os controles. Houve efeito significativo da dieta e da infecção no ganho de peso (P 0,05). O ganho em peso diário foi de 0,170 kg no G1, 0,205 kg no G2, 0,085 kg no G3 e 0,116 kg no G4. Os grupos infectados, apesar de receberem tratamentos anti-helmínticos preventivos que evitaram mortes por haemonchose, apresentaram redução nos parâmetros produtivos em comparação com os controles.(AU)

Helminth infections and hybridization between Haemonchus contortus and Haemonchus placei in sheep from Santana do Livramento, Brazil / Infecções helmínticas e hibridização entre Haemonchus contortus e Haemonchus placei em ovinos criados em Santana do Livramento, Brasil

The occurrence and intensity of helminth infections were evaluated in sheep from pastures shared with cattle. In 2015 and 2016, young male sheep acquired in Santana do Livramento, Rio Grande do Sul, Brazil, were finished in integrated crop-livestock system. We selected the 12 sheep that showed the highest number of nematode eggs per gram of faeces to search for worms in the gastrointestinal tract. Haemonchus contortus and Trichostrongylus colubriformis were the major parasites. H. contortus presented mean intensities of 1,159 and 257 worms in 2015 and 2016, respectively. T. colubriformis displayed mean intensities of 4,149 and 2,427 worms in 2015 and 2016, respectively. Of the 127 male specimens of Haemonchus spp. analysed by Polymerase Chain Reaction (PCR), 125 were H. contortus, one Haemonchus placei and one hybrid. Other species detected were Cooperia punctata , Cooperia pectinata, Cooperia spatulata, Cooperia curticei, Ostertagia ostertagi, Teladorsagia circumcincta , Trichostrongylus axei, Nematodirus spathiger , and Trichuris ovis. Twenty lambs presented cysts of Taenia hydatigena in the liver and mesentery. One lamb presented Coenurus cerebralis, the larval stage of Taenia multiceps, in the brain. In conclusion, sheep from pasture shared with cattle presented a high diversity of nematode species. H. contortus and H. placei co-infection occur with consequent hybridization.(AU)

A ocorrência de infecções helmínticas foi avaliada em ovinos que compartilhavam pastagem com bovinos. Em 2015 e em 2016, cordeiros machos foram adquiridos em Santana do Livramento, Rio Grande do Sul, para serem terminados em sistema de lavoura pecuária (ILP). Em cada ano, 12 cordeiros que tinham maior contagem de ovos nas fezes foram abatidos para recuperação dos vermes. Haemonchus contortus e Trichostrongylus colubriformis foram as principais espécies registradas. H. contortus apresentou intensidade média de 1159 e 257 vermes em 2015 e 2016, respectivamente. T. colubriformis apresentou intensidade média de 4149 e 2427 parasitas em 2015 e 2016, respectivamente. De 127 machos de Haemonchus spp. analisados por Polymerase Chain Reaction (PCR), 125 foram identificados como H. contortus, um como Haemonchus placei e um como híbrido. Cooperia punctata, Cooperia pectinata, Cooperia spatulata, Cooperia curticei, Ostertagia ostertagi, Teladorsagia circumcincta , Trichostrongylus axei, Nematodirus spathiger e Trichuris ovis foram as outras espécies de nematódeos registradas. Vinte cordeiros apresentaram cistos de Taenia hydatigena no mesentério e no fígado. Um cordeiro apresentou no cérebro Coenurus cerebralis, o estágio larval de Taenia multiceps . Em conclusão, ovinos criados com bovinos apresentam grande diversidade de nematódeos. A co-infecção de H. contortus e H. placei favorece a produção de híbridos.(AU)

Helminth infections and hybridization between Haemonchus contortus and Haemonchus placei in sheep from Santana do Livramento, Brazil / Infecções helmínticas e hibridização entre Haemonchus contortus e Haemonchus placei em ovinos criados em Santana do Livramento, Brasil

Abstract The occurrence and intensity of helminth infections were evaluated in sheep from pastures shared with cattle. In 2015 and 2016, young male sheep acquired in Santana do Livramento, Rio Grande do Sul, Brazil, were finished in integrated crop-livestock system. We selected the 12 sheep that showed the highest number of nematode eggs per gram of faeces to search for worms in the gastrointestinal tract. Haemonchus contortus and Trichostrongylus colubriformis were the major parasites. H. contortus presented mean intensities of 1,159 and 257 worms in 2015 and 2016, respectively. T. colubriformis displayed mean intensities of 4,149 and 2,427 worms in 2015 and 2016, respectively. Of the 127 male specimens of Haemonchus spp. analysed by Polymerase Chain Reaction (PCR), 125 were H. contortus, one Haemonchus placei and one hybrid. Other species detected were Cooperia punctata , Cooperia pectinata, Cooperia spatulata, Cooperia curticei, Ostertagia ostertagi, Teladorsagia circumcincta , Trichostrongylus axei, Nematodirus spathiger , and Trichuris ovis. Twenty lambs presented cysts of Taenia hydatigena in the liver and mesentery. One lamb presented Coenurus cerebralis, the larval stage of Taenia multiceps, in the brain. In conclusion, sheep from pasture shared with cattle presented a high diversity of nematode species. H. contortus and H. placei co-infection occur with consequent hybridization.

Resumo A ocorrência de infecções helmínticas foi avaliada em ovinos que compartilhavam pastagem com bovinos. Em 2015 e em 2016, cordeiros machos foram adquiridos em Santana do Livramento, Rio Grande do Sul, para serem terminados em sistema de lavoura - pecuária (ILP). Em cada ano, 12 cordeiros que tinham maior contagem de ovos nas fezes foram abatidos para recuperação dos vermes. Haemonchus contortus e Trichostrongylus colubriformis foram as principais espécies registradas. H. contortus apresentou intensidade média de 1159 e 257 vermes em 2015 e 2016, respectivamente. T. colubriformis apresentou intensidade média de 4149 e 2427 parasitas em 2015 e 2016, respectivamente. De 127 machos de Haemonchus spp. analisados por "Polymerase Chain Reaction" (PCR), 125 foram identificados como H. contortus, um como Haemonchus placei e um como híbrido. Cooperia punctata, Cooperia pectinata, Cooperia spatulata, Cooperia curticei, Ostertagia ostertagi, Teladorsagia circumcincta , Trichostrongylus axei, Nematodirus spathiger e Trichuris ovis foram as outras espécies de nematódeos registradas. Vinte cordeiros apresentaram cistos de Taenia hydatigena no mesentério e no fígado. Um cordeiro apresentou no cérebro Coenurus cerebralis, o estágio larval de Taenia multiceps . Em conclusão, ovinos criados com bovinos apresentam grande diversidade de nematódeos. A co-infecção de H. contortus e H. placei favorece a produção de híbridos.

Immune and haematological parameters of Blackbelly ewes infected with gastrointestinal nematodes / Parámetros inmunológicos y hematológicos en ovejas Blackbelly infectadas con nematodos gastrointestinales / Parâmetros imunes e hematológicos de ovelhas Blackbelly infectadas por nematoides gastrintestinais

Abstract Background: It is necessary to identify phenotypic traits related to natural resistance against gastrointestinal nematodes (GIN) in order to know the host immunity status in productive ewes. Objetive: To determine haematological and immunological parameters (IgA and IgG) during pregnancy and lactation in Blackbelly ewes naturally infected with GIN Methods: The number of eggs per gram (EPG), packed cell volume (% PCV), plasmatic protein (PP), and peripheral eosinophils were determined during eight months. In addition, sera and saliva samples were collected to establish IgG and IgA kinetics by indirect enzyme-linked immunosorbent assay (ELISA). Results: The results showed 2,592 ± 2,403 EPG and 22.2 ± 4.0% PCV during lactation and 595 ± 901 EPG and 25.1 ± 2.5% PCV during pregnancy. A higher percentage of Trichostrongylus colubriformis larvae were observed in pregnancy (84 to100%) than in lactation (36 to 44%). The IgA activity in serum samples showed a marked reduction (from 80 to 10%) during lambing for both Haemonchus contortus and T. colubriformis antigens. In saliva samples, IgA activity with regard to the standard decreased from 56% at 60 days to 30% at 45 days before lambing and remained low for 45 days during lactation (23 to 32% activity). The eosinophils numbers were 2.0 x 109 cells L-1 in pregnancy and remained low at 0.7 x 109 cells L-1 in lactation. Conclusion: The studied variables reflect the breakdown of immunity against GIN in Blackbelly ewes before and after lambing.

Resumen Antecedentes: La identificación del fenotipo relacionado con la resistencia contra nematodos gastrointestinales (GIN) es necesaria para conocer la inmunidad del huésped en ovejas en producción. Objetivo: Determinar los parámetros hematológicos e inmunológicos (IgA e IgG) en gestación y lactancia en ovejas Blackbelly infectadas naturalmente con GIN. Métodos: Se determinó el número de huevos por gramo de heces (EPG), se identificaron las larvas y se registró el porcentaje del volumen celular aglomerado (% PCV), proteína plasmática (PP) y eosinófilos periféricos durante ocho meses. Además, se colectó suero y saliva para determinar la cinética de IgG e IgA por medio de un ensayo inmuno-enzimático (ELISA) indirecto. Resultados: Los resultados mostraron 2.592 ± 2.403 EPG y 22,2 ± 4,0% PCV durante la lactancia y 595 ± 901 EPG y 25,1 ± 2,5% PCV durante la gestación. Se observó un mayor porcentaje de larvas de Trichostrongylus colubriformis en gestación (84 a 100%) que en lactancia (36 a 44%). La actividad de la IgA en las muestras de suero mostró una marcada reducción después del parto para ambos antígenos de Haemonchus contortus y T. colubriformis (80 a 10%). En saliva, la actividad de la IgA disminuyó de 56 a 30% del día 60 al 45 antes del parto y se mantuvo baja en los primeros 45 días de la lactancia (actividad de 23 a 32%). El número de eosinófilos fue de 2,0 x 109 células L-1 durante la gestación, y se redujo a 0,7 x 109 células L-1 en la lactancia. Conclusión: Las variables estudiadas reflejan la ruptura de la inmunidad contra GIN en ovejas Blackbelly antes y después del parto.

Resumo Antecedentes: A identificação de traços fenotípicos relacionados à resistência natural contra nematóides gastrintestinais (GIN) é necessária para saber a imunidade do hospedeiro em ovelhas produtivas. Objetivo: Determinar parâmetros hematológicos e imunológicos (IgA e IgG) em períodos de gestação e lactação em ovelhas Blackbelly naturalmente infectados com GIN. Métodos: O número de ovos por grama (EPG), volume empacotado de células (% PCV), proteína plasmática (PP) e eosinófilos periféricos foram determinados durante oito meses. Além disso, as amostras de soro e saliva foram recolhidas para determinar a cinética de IgG e IgA por ELISA indireto. Resultados: Os resultados mostraram 2.592 ± 2.403 EPG e 22,2 ± 4,0% PCV durante a lactação e 595 ± 901 EPG e 25,1 ± 2,5% PCV durante a gravidez. A percentagem mais elevada de larvas de Trichostrongylus colubriformis foi observada na gravidez (84 a 100%) do que na lactação (36 a 44%). A atividade de IgA em amostras de soro mostrou uma redução acentuada (80 a 10%) durante o parto nos antígenos de Haemonchus contortus e T. colubriformis. Em amostras de saliva, a atividade de IgA diminuiu de 56 a 30% do dia 60 ao 45 antes do parto e permaneceu baixa por 45 dias durante a lactação (atividade de 23 a 32%). O número de eosinófilos foi de 2,0 x 109 células L-1 na gravidez e manteve-se baixo, com 0,7 x 109 células L-1 na lactação. Conclusaõ: As variáveis estudadas refletem a quebra da imunidade contra GIN em ovelhas Blackbelly antes e depois do parto.

Sobre algunos helmintos parásitos de la taruca, Hippocamelus antisensis (Mammalia: Artiodactyla) / On some helminth parasites of the taruca, Hippocamelus antisensis (Mammalia: Artiodactyla)

En el presente estudio se da a conocer tres especies de helmintos parásitos colectados de una taruca (Hippocamelus antisensis) en el Perú. Una taruca macho adulto fue remitido al Instituto Veterinario de Investigaciones Tropicales y de Altura (IVITA), Sede Maranganí, por el Servicio Nacional Forestal y de Fauna Silvestre del Perú (SERFOR), para su respectiva necropsia. Se colectaron 2 nematodos del sistema digestivo, así como un quiste localizado en el omento. Para el diagnóstico parasitológico se evaluaron las características morfo-métricas de los especímenes colectados. Los nematodos fueron diagnosticados como Trichostrongylus axei y Mazamastrongylus sp.; por otro lado, el quiste fue diagnosticado como un metacestodo de Taenia hydatigena. El hallazgo de T. axei representa el primer registro para la taruca. En la presente nota se realiza también una breve descripción morfológica de los helmintos mencionados.

This study presents three species of helminth parasites collected from a taruca (Hippocamelus antisensis) in Peru. Three helminth parasites from the taruca (Hippocamelus antisensis) are described in this report. An adult male taruca was remitted to the Veterinary Institute of Tropical and Highland Research (IVITA), Sede Maranganí, for the National Forest and Wildlife Service of Peru (SERFOR) for their respective necropsy. Two nematodes were collected from the digestive system, and 1 cyst was collected from the omentum. Morphometric characteristics of the specimens were evaluated for the parasitological diagnosis. Nematodes were diagnosed as Trichostrongylus axei and Mazamastrongylus sp.; on the other hand, the cyst was diagnosed as a metacestode of Taenia hydatigena. T. axei finding represents the first record for the taruca. A brief morphological description of each helminth and discussion about their hosts and distribution is done.

Dried, ground banana plant leaves (Musa spp.) for the control of Haemonchus contortus and Trichostrongylus colubriformis infections in sheep.

To evaluate the anthelmintic effect of Musa spp. leaves, 12 animals were artificially infected with Haemonchus contortus, and another 12 animals were infected with Trichostrongylus colubriformis. Then, both treatment groups were offered 400 g of dried ground banana plant leaves, and the control animals were offered only 1000 g of coast cross hay. During the trials, the animals received weekly physical examinations. The methods used to evaluate the efficiency of this treatment were packed cell volume, total plasma protein and faecal egg counts, and egg hatchability tests were performed on days -2, +3, +6, +9, +13 and +15. Coproculture tests were performed on day -2 to confirm monospecific infections. In the FEC and EHT, a statistically significant difference (0.04, 0.005; p < 0.05) was noted for T. colubriformis. There were no statistically significant differences (p > 0.05) for Haemochus contortus group in all tests. Our results confirmed previous findings suggesting that dried ground banana plant leaves possess anthelmintic activity.

Gastrointestinal nematode infections in sheep raised in Botucatu, state of São Paulo, Brazil / Infecções por nematódeos gastrintestinais em ovinos criados em Botucatu, estado de São Paulo, Brasil

Gastrointestinal nematode infections were evaluated in sheep raised in Botucatu, state of São Paulo, Brazil between April 2008 and March 2011. Every month, two tracer lambs grazing with a flock of sheep were exposed to natural infection with gastrointestinal nematodes for 28 consecutive days. At the end of this period, the lambs were sacrificed for worm counts. Haemonchus contortus presented 100% of prevalence. The seasons exerted no significant influence on the mean intensity of H. contortus, which ranged from 315 worms in November 2010 to 2,5205 worms in January 2011. The prevalence of Trichostrongylus colubriformis was also 100%, with the lowest mean intensity (15 worms) recorded in February 2011 and the highest (9,760 worms) in October 2009. In the case of T. colubriformis, a significant correlation coefficient was found between worm counts vs. rainfall (r = −0.32; P <0.05). Three other nematodes species were found in tracer lambs, albeit in small numbers. Their prevalence and mean intensity (in parenthesis) were as follows: Oesophagostomum columbianum 28% (25.2), Cooperia curticei 7% (4.5) and Trichuris spp. 2% (1). In conclusion, the environmental conditions of the area proved to be highly favorable for the year-round transmission of H. contortus and T. colubriformis.

A ocorrência de infecções por nematódeos gastrintestinais foi avaliada de abril de 2008 até março de 2011em ovinos criados em Botucatu, estado de São Paulo. Todos os meses, dois cordeiros traçadores foram expostos à infecção natural por nematódeos gastrintestinais, durante 28 dias consecutivos, ao pastejar junto com um rebanho de ovelhas. Ao final desse período, os animais foram sacrificados para a identificação e quantificação dos helmintos. Haemonchus contortus apresentou prevalência de 100%. Não houve influência significativa das estações do ano na intensidade média de H. contortus, que variou de 315 vermes em novembro/2010 a 25.205 vermes em janeiro/2011. Trichostrongylus colubriformis também apresentou prevalência de 100% com a menor intensidade média (15 vermes) em fevereiro/2011 e a maior (9.760 vermes) em outubro/2009. No caso de T. colubriformis, houve correlação significativa entre as contagens de vermes x precipitação (r = −0,32; P <0,05). Outras três espécies de nematódeos foram encontradas nos cordeiros traçadores, no entanto em pequenas quantidades, com as seguintes prevalências e intensidades médias (entre parênteses): Oesophagostomum columbianum 28% (25,2), Cooperia curticei 7% (4,5) e Trichuris spp. 2 % (1). Em conclusão, as condições ambientais da área foram muito favoráveis durante todo o ano para a transmissão de H. contortus e T. colubriformis.

Gastrointestinal nematode infections in sheep raised in Botucatu, state of São Paulo, Brazil / Infecções por nematódeos gastrintestinais em ovinos criados em Botucatu, estado de São Paulo, Brasil

Gastrointestinal nematode infections were evaluated in sheep raised in Botucatu, state of São Paulo, Brazil between April 2008 and March 2011. Every month, two tracer lambs grazing with a flock of sheep were exposed to natural infection with gastrointestinal nematodes for 28 consecutive days. At the end of this period, the lambs were sacrificed for worm counts. Haemonchus contortus presented 100% of prevalence. The seasons exerted no significant influence on the mean intensity of H. contortus, which ranged from 315 worms in November 2010 to 2,5205 worms in January 2011. The prevalence of Trichostrongylus colubriformis was also 100%, with the lowest mean intensity (15 worms) recorded in February 2011 and the highest (9,760 worms) in October 2009. In the case of T. colubriformis, a significant correlation coefficient was found between worm counts vs. rainfall (r = −0.32; P <0.05). Three other nematodes species were found in tracer lambs, albeit in small numbers. Their prevalence and mean intensity (in parenthesis) were as follows: Oesophagostomum columbianum 28% (25.2), Cooperia curticei 7% (4.5) and Trichuris spp. 2% (1). In conclusion, the environmental conditions of the area proved to be highly favorable for the year-round transmission of H. contortus and T. colubriformis.

A ocorrência de infecções por nematódeos gastrintestinais foi avaliada de abril de 2008 até março de 2011em ovinos criados em Botucatu, estado de São Paulo. Todos os meses, dois cordeiros traçadores foram expostos à infecção natural por nematódeos gastrintestinais, durante 28 dias consecutivos, ao pastejar junto com um rebanho de ovelhas. Ao final desse período, os animais foram sacrificados para a identificação e quantificação dos helmintos. Haemonchus contortus apresentou prevalência de 100%. Não houve influência significativa das estações do ano na intensidade média de H. contortus, que variou de 315 vermes em novembro/2010 a 25.205 vermes em janeiro/2011. Trichostrongylus colubriformis também apresentou prevalência de 100% com a menor intensidade média (15 vermes) em fevereiro/2011 e a maior (9.760 vermes) em outubro/2009. No caso de T. colubriformis, houve correlação significativa entre as contagens de vermes x precipitação (r = −0,32; P <0,05). Outras três espécies de nematódeos foram encontradas nos cordeiros traçadores, no entanto em pequenas quantidades, com as seguintes prevalências e intensidades médias (entre parênteses): Oesophagostomum columbianum 28% (25,2), Cooperia curticei 7% (4,5) e Trichuris spp. 2 % (1). Em conclusão, as condições ambientais da área foram muito favoráveis durante todo o ano para a transmissão de H. contortus e T. colubriformis.

Fifteen years later, anthelmintic resistances have dramatically spread over goat farms in Guadeloupe.

Faecal egg count reduction tests (FECRTs) were performed on 21 goat farms in Guadeloupe (FWI). Anthelmintic resistance (AR) to netobimin (benzimidazole) was found in all 15 herds in which it was tested. AR to ivermectin (avermectin) and levamisole (imidazothiazole) were also very largely spread (14 out of 17 farms and 7 out of 9 farms, respectively). AR to the final moxidectin (milbemycin) released was already present in 2 out of 9 farms in which it was tested. Haemonchus was the dominant genus of gastrointestinal nematodes and was more frequently found to be resistant to netobimin, ivermectin and moxidectin than Trichostrongylus, the latter appeared to be more often resistant to levamisole. A first survey 15 years ago revealed only AR to benzimidazoles and one suspected case of AR to ivermectin.

Gastrointestinal nematodiosis of goats: a review / Nematodioses gastrointestinais de caprinos: uma revisão

The gastrointestinal nematodiosis represents one of the main health problems of goats reared under grazing, and in Haemonchus contortus and Trichostrongylus colubriformis species it is the most responsible for economic losses of goat. The diagnosis of parasitic animal must be grounded in clinical evaluations and results of tests used, including the pasture. The degree of parasitic infection in animal and pasture contamination assist in the adoption of measures of grazing management (forage, height of grazing, pasture rotation and stocking rate) and anthelmintic treatment suitable for each system created in different geographic regions. This article was written according to the health and economic importance of gastrointestinal helminthiasis caused by nematodes in goats subjected to grazing.(AU)

As nematodioses gastrintestinais representam um dos principais problemas sanitários de caprinos criados em sistema de pastejo, sendo as espécies Haemonchus contortus e Trichostrongylus colubriformis os maiores responsáveis pelos prejuízos econômicos da caprinocultura. O diagnóstico do parasitismo animal deve ser embasado em avaliações clínicas e nos resultados dos exames parasitológicos empregados, inclusive na pastagem. O grau de infecção parasitária nos animais e a contaminação da pastagem auxiliam na adoção de medidas de manejo do pastejo (altura do pastejo, espécie forrageira, rotação de pastagem, taxa de lotação) e no tratamento anti-helmíntico adequados a cada sistema de criação, em diferentes regiões geográficas. De acordo com a importância sanitária e econômica das helmintoses gastrintestinais causadas por nematódeos em caprinos submetidos ao pastejo foi elaborado este artigo de revisão.(AU)

Gastrointestinal nematodiosis of goats: a review / Nematodioses gastrointestinais de caprinos: uma revisão

The gastrointestinal nematodiosis represents one of the main health problems of goats reared under grazing, and in Haemonchus contortus and Trichostrongylus colubriformis species it is the most responsible for economic losses of goat. The diagnosis of parasitic animal must be grounded in clinical evaluations and results of tests used, including the pasture. The degree of parasitic infection in animal and pasture contamination assist in the adoption of measures of grazing management (forage, height of grazing, pasture rotation and stocking rate) and anthelmintic treatment suitable for each system created in different geographic regions. This article was written according to the health and economic importance of gastrointestinal helminthiasis caused by nematodes in goats subjected to grazing.

As nematodioses gastrintestinais representam um dos principais problemas sanitários de caprinos criados em sistema de pastejo, sendo as espécies Haemonchus contortus e Trichostrongylus colubriformis os maiores responsáveis pelos prejuízos econômicos da caprinocultura. O diagnóstico do parasitismo animal deve ser embasado em avaliações clínicas e nos resultados dos exames parasitológicos empregados, inclusive na pastagem. O grau de infecção parasitária nos animais e a contaminação da pastagem auxiliam na adoção de medidas de manejo do pastejo (altura do pastejo, espécie forrageira, rotação de pastagem, taxa de lotação) e no tratamento anti-helmíntico adequados a cada sistema de criação, em diferentes regiões geográficas. De acordo com a importância sanitária e econômica das helmintoses gastrintestinais causadas por nematódeos em caprinos submetidos ao pastejo foi elaborado este artigo de revisão.

Human infection by Trichostrongylus spp. in residents of urban areas of Salvador city, Bahia, Brazil / Infección humana por Trichostrongylus spp. en residentes de zonas urbanas de la ciudad de Salvador, Bahia, Brasil

Introduction: Nematodes of Trichostrongylus genus are mainly parasites of herbivores, although sporadic human infections have been reported in many countries. Objective: To describe the frequency and seasonality of Trichostrongylus spp. infection in individuals attended at a public clinical laboratory. Materials and methods: Fecal samples of 9,283 individuals were evaluated by spontaneous sedimentation (Lutz) in the Parasitology Laboratory of the Pharmacy College, Federal University of Bahia, Brazil, from January of 2006 to May of 2008. The positive samples for either Trichostrongylus spp. or hookworms were further examined to evaluate the morphometry of nematode eggs. Results: One-hundred and ten patients (1.2%) were confirmed to be infected by Trichostrongylus spp. The positive cases were significantly more frequent in females (1.6%; p <0.05), with higher distribution in the age group between 11-20 years (1.9%), compared to those aged 51-60 (0.8%) and older than 60 years (0.9%)( p <0.05), independent of gender. Trichostrongylus spp. infections were more common from March to May (40 cases) and showed a homogeneous distribution over the other periods of the year (21-25 cases). The hematological analyses of 60 Trichostrongylus -infected patients showed normal levels of eosinophils in most of the positive cases. Conclusions: The data reveal that the occurrence of infection by Trichostrongylus spp. in residents of Salvador is more frequent than those reported in other urban regions and that it is essential to distinguish the parasite from other nematodes in routine parasitological examination.

Introducción. Los nematodos del género Trichostrongylus son parásitos principalmente herbívoros, aunque se hayan descrito en muchos países infecciones humanas esporádicas. Objetivo. Describir la frecuencia y estacionalidad de la infección por Trichostrongylus spp. en individuos atendidos en un laboratorio clínico público. Materiales y métodos. Las muestras fecales de 9.283 individuos fueron analizadas mediante sedimentación espontánea (Lutz) en el Laboratorio de Parasitología de la Facultad de Farmacia de la Universidad Federal de Bahía, Brasil, desde enero de 2006 a mayo de 2008. Las muestras positivas para Trichostrongylus o Ancylostoma fueron reexaminadas para evaluar la morfometría de los huevos de nematodos. Resultados. La infección por Trichostrongylus spp. se confirmó en 110 pacientes (1,2 %), con mayor frecuencia en el sexo femenino (1,6 %; p<0,05). La frecuencia fue mayor en el grupo de edad de 11 a 20 años (1,9 %), independientemente del sexo, en comparación con las personas de 51 a 60 (0,8 %) y con los mayores de 60 años (0,9 %) (p < 0,05). Las infecciones por Trichostrongylus spp. fueron más frecuentes entre marzo y mayo (40 casos) con una distribución homogénea en otros periodos del año (21 a 25 casos). Los análisis hematológicos de 60 pacientes infectados con Trichostrongylus spp. presentaron niveles normales de eosinófilos en la mayoría de los casos positivos. Conclusiones. Los datos revelan que la incidencia de la infección por Trichostrongylus spp. en individuos residentes en Salvador es más frecuente que las descritas en otras áreas urbanas y, por ello, es esencial distinguir dicho parásito de otros nematodos en los exámenes parasitológicos de rutina.