Reduction of egg reappearance period of cyathostomins in naturally infected horses after increasing doses of ivermectin in Brazil: a grim picture for sustainable parasite control.

Cyathostomins are the largest group of parasites in horses that can be controlled by ivermectin (IVM). This study aimed to run a four-dose titration trial of IVM in 28 naturally infected Thoroughbred yearlings. The local Strongyle population had been recorded to be resistant to IVM (200 µg/kg). The parasite fecal egg count (FEC) was performed to investigate the egg reappearance period (ERP) of two and five weeks (w2pt and w5pt) after IVM treatment. FEC was > 1000 on day zero for all groups. Although 100% FEC reduction was reported at w2pt for all concentrations, the FEC at w5pt revealed < 83% efficacy. This study reports the reduction of ERP using the label dose as well as 300, and 400 µg/kg (double dose) of IVM. The protocol allowed IVM to significantly suppress FEC w2pt although not eliminating adult worms, failing to guarantee an extension of its protection period over 8 weeks. Moreover, the FEC at w5pt possibly means the infection was not cleared, and worms reestablished egg laying. We raised the possibility of withdrawing IVM of control programs when the drug has less than 80% FEC reduction at w5pt.

Prevalence and risk factors of gastrointestinal helminths infection in Brazilian horses: A retrospective study of a 12-year (2008-2019) diagnostic data.

Understanding gastrointestinal parasite distribution is crucial for effective control programs in horses. This study reports the prevalence of helminth infections in horses and selected risk factors (i.e., breed, age, climate, season) by analyzing 19,276 fecal samples from the Laboratory of Veterinary Clinical Parasitology, in Curitiba, Southern Brazil. The analyses were carried out from 2008 to 2019, coming from 153 stud farms located in 60 municipalities of nine Brazilian states. The parasite prevalence was 73.3%, with 72.1% present in the adult population and 80.6% in young horses. Strongyles were present in 100% horse farms. Strongyles had a prevalence of 72.1% with a mean FEC of 453.53 (+/- 717.6). Parascaris spp. had a prevalence of 5.8% and a FEC of 17.11 (+/- 149.2). The tropical wet/monsoon climate (Am) showed the lowest FEC for strongyles and Parascaris spp. when compared to the other climates. In the logistic regression analysis, young horses exhibited 4.6 times higher odds ratio (OR) (3.9-5.5) of Parascaris spp. and 1.2 (1.1-1.4) times higher OR of strongyles egg shedding when compared to adults (P < 0.001). Summer presented a higher risk for Parascaris spp. and Strongyles eggs when compared to the other seasons (P < 0.001). Mangalarga Marchador, Criollo, and Crossbred breeds were identified with higher OR of Parascaris spp. egg shedding than Thoroughbred. The extensive prevalence of strongyles across ages, seasons, breeds, and climates alerts for the risk of clinical manifestations in equines raised on pastures designing optimal health management and parasite control strategies worldwide.

Acaricidal effect of major compounds to control Rhipicephalus microplus (Canestrini, 1887) in dairy cows and possible alternatives for reversing multidrug resistance.

Tick control represent a great challenge to animal health. The objective of this study was to evaluate the efficacy of acaricidal compounds against Rhipicephalus microplus from dairy cattle systems in Paraná State, Brazil. Six farms (PR1-PR6) were selected, where anti-tick products were applied at fixed intervals. Two other farms that adopted alternative protocols; target selective treatment (PRS), and individual-based agroecological protocol (PRA) were also included in the trial. Fully engorged R. microplus were collected for the in vitro adult immersion test (AIT), and the egg hatch test (EHT) in all eight populations. The larval packet test (LPT) was used on PR6 and PRA tick populations. The treatment groups were composed of G1: dichlorvos 45% plus cypermethrin 5%, G2: deltamethrin 2.5%, G3: cypermethrin 15%, chlorpyriphos 25%, plus citronellal 1%, and G4: amitraz 12.5%. The efficacy at PR1 to PR6 revealed that G3 and G4 achieved moderate to high efficacy, from 75.0 to 100.0% and 73 to 98%, respectively. In the LPT, the efficacy at PR6 was 76.0, 67.0, 93.0 and 30.6%, while PRA presented 100.0, 100.0, 100.0, and 54.0%, for G1, G2, G3 and G4, respectively. Sustainable parasite control strategies are discussed.

Acaricidal effect of major compounds to control Rhipicephalus microplus (Canestrini, 1887) in dairy cows and possible alternatives for reversing multidrug resistance / Efeito acaricida dos principais compostos no controle de Rhipicephalus microplus (Canestrini, 1887) em vacas leiteiras e possíveis alternativas para reverter a resistência múltipla às drogas

Tick control represent a great challenge to animal health. The objective of this study was to evaluate the efficacy of acaricidal compounds against Rhipicephalus microplus from dairy cattle systems in Paraná State, Brazil. Six farms (PR1-PR6) were selected, where anti-tick products were applied at fixed intervals. Two other farms that adopted alternative protocols; target selective treatment (PRS), and individual-based agroecological protocol (PRA) were also included in the trial. Fully engorged R. microplus were collected for the in vitro adult immersion test (AIT), and the egg hatch test (EHT) in all eight populations. The larval packet test (LPT) was used on PR6 and PRA tick populations. The treatment groups were composed of G1: dichlorvos 45% plus cypermethrin 5%, G2: deltamethrin 2.5%, G3: cypermethrin 15%, chlorpyriphos 25%, plus citronellal 1%, and G4: amitraz 12.5%. The efficacy at PR1 to PR6 revealed that G3 and G4 achieved moderate to high efficacy, from 75.0 to 100.0% and 73 to 98%, respectively. In the LPT, the efficacy at PR6 was 76.0, 67.0, 93.0 and 30.6%, while PRA presented 100.0, 100.0, 100.0, and 54.0%, for G1, G2, G3 and G4, respectively. Sustainable parasite control strategies are discussed.(AU)

O controle do carrapato representa um grande desafio na saúde animal. O objetivo deste trabalho foi avaliar a eficácia de compostos acaricidas contra Rhipicephalus microplus em sistemas de pecuária leiteira no Paraná, Brasil. Foram selecionadas seis fazendas (PR1-PR6), onde são aplicados produtos carrapaticidas regularmente. Duas outras fazendas que adotaram protocolos alternativos, PRS (tratamento seletivo) e PRA (controle agroecológico individual), também foram incluídas nos testes. R. microplus ingurgitadas foram coletados para testes in vitro de imersão em adultos (TIA) e teste de eclosão de ovos (TEO) em todas as oito populações. O teste de envelope de larvas (TEL) foi utilizado nas populações PR6 e PRA. Os grupos de tratamento foram compostos por G1: diclorvos 45% mais cipermetrina 5%, G2: deltametrina 2,5%, G3: cipermetrina 15%, clorpirifos 25% e mais citronelal 1% e G4: amitraz 12,5%. A eficácia em PR1 a PR6 revelou que G3 e G4 obtiveram eficácia de moderada a alta, variando de 75,0% a 100,0% e 73% a 98%, respectivamente. No TEL, a eficácia em PR6 foi de 76,0%, 67,0%, 93,0% e 30,6%, enquanto em PRA apresentou 100,0%, 100,0%, 100,0% e 54,0%, para G1, G2, G3 e G4, respectivamente. Estratégias sustentáveis de controle são discutidas neste documento.(AU)

Occurrence of gastrointestinal parasites in wild animals in State of Paraná, Brazil

ABSTRACT The objective of this study was to determine the prevalence and diversity of gastrointestinal parasites in fecal samples from wild birds and mammals from the State of Paraná. In total, 220 stool samples were sent to Parasitic Diseases Laboratory of the Federal University of Paraná during 13 months (Jan/2013-Jan/2014). A total of 52.7% (116/220) of the animals were positive for cysts, oocysts, eggs and/or trophozoites. In birds, the positivity rate was 37.9% (25/66) and mammals was 59.1% (91/154). Strongyloidea superfamily eggs were observed in 37.3% (82/220) of the samples, Eimeria spp. in 10% (22/220), and Trichuris spp. in 4.5% (10/220). The most frequent mammal species were llamas (Lama glama), and dromedaries (Camelus bactrianus) with infection rate of 70.1% (54/77) and 60.8% (14/23), respectively. In other hand, cockatiels (Nymphicus hollandicus) and ring necks (Psittacula krameri), were the most researched birds, with infection rate of 20% (40/50) and 100% (6/6), respectively. A high prevalence of gastrointestinal parasites was observed in most of wildlife animals. Further investigations should be conducted focusing on parasite control strategies and the conservation measurements for harmonizing the human-animal interaction on the long-term, reducing associated health risks.

Occurrence of gastrointestinal parasites in wild animals in State of Paraná, Brazil.

The objective of this study was to determine the prevalence and diversity of gastrointestinal parasites in fecal samples from wild birds and mammals from the State of Paraná. In total, 220 stool samples were sent to Parasitic Diseases Laboratory of the Federal University of Paraná during 13 months (Jan/2013-Jan/2014). A total of 52.7% (116/220) of the animals were positive for cysts, oocysts, eggs and/or trophozoites. In birds, the positivity rate was 37.9% (25/66) and mammals was 59.1% (91/154). Strongyloidea superfamily eggs were observed in 37.3% (82/220) of the samples, Eimeria spp. in 10% (22/220), and Trichuris spp. in 4.5% (10/220). The most frequent mammal species were llamas (Lama glama), and dromedaries (Camelus bactrianus) with infection rate of 70.1% (54/77) and 60.8% (14/23), respectively. In other hand, cockatiels (Nymphicus hollandicus) and ring necks (Psittacula krameri), were the most researched birds, with infection rate of 20% (40/50) and 100% (6/6), respectively. A high prevalence of gastrointestinal parasites was observed in most of wildlife animals. Further investigations should be conducted focusing on parasite control strategies and the conservation measurements for harmonizing the human-animal interaction on the long-term, reducing associated health risks.

Diagnosis of resistance alleles in codon 167 of the beta-tubulin (Cya-tbb-1) gene from third-stage larvae of horse cyathostomins.

Anthelmintic resistance is a serious problem for the control of equine gastrointestinal nematodes. In the present survey, 173 third stage larvae of cyathostomins were investigated from three different locations for the presence of the resistant genotype at codon 167 of the beta-tubulin gene, as this is the most prevalent mutation. The larvae from the state of Parana (n=67), Sao Paulo (n=54) and Santa Catarina (n=52), showed 61.2; 31.5 and 38.5% of the heterozygous resistant genotype - TTC/TAC, respectively. An unpublished mutation at codon 172 that results in a serine (S) to threonine (T) substitution was found in 17.9% (12/67) of samples from Parana; and 13.0% (7/54) of samples from Sao Paulo. We have compared the molecular diagnostic with the fecal egg count data (R2=-0.79) from the same farms, and consider that the use of routine molecular diagnostic in individual larva may help to determine the population genetic distribution that is associated with drug failure.

Evaluation of resistance in a selected field strain of Haemonchus contortus to ivermectin and moxidectin using the Larval Migration on Agar Test / Avaliação da resistência em um isolado de campo selecionado de Haemonchus contortus à ivermectina e moxidectina usando o Teste de Migração de Larvas em Ágar

Haemonchus contortus is one of the most common and economically significant causes of disease in small ruminants worldwide, and the control programs of parasitic nematodes - including H. contortus - rely mostly on the use of anthelmintic drugs. The consequence of the use of this, as the sole sanitary strategy to avoid parasite infections, was the reduction of the efficacy of all chemotherapeutic products with a heavy selection for resistance. The widespread of anthelmintic resistance and the difficulty of its early diagnosis has been a major concern for the sustainable parasite management on farms. The objective of this research was to determine and compare the ivermectin (IVM) and moxidectin (MOX) effect in a selected field strain of H. contortus with a known resistance status, using the in vitro larval migration on agar test (LMAT). Third stage larvae of the selected isolate were obtained from faecal cultures of experimentally infected sheep and incubated in eleven increasing diluted concentrations of IVM and MOX (6, 12, 24, 48, 96, 192, 384, 768, 1536, 3072 and 6144µg/mL). The dose-response sigmoidal curves were obtained using the R² value of >0.90 and the lethal concentration (LC50) dose for the tested anthelmintic drugs using a four-parameter logistic model. The LC50 value for MOX was significantly lower than IVM (1.253µg/mL and 91.06µg/mL), identifying the H. contortus isolate as considerably less susceptible to IVM compared to MOX. Furthermore, the LMAT showed a high consistency (p<0.0001) and provided to be a useful diagnostic tool for monitoring the resistance status of IVM and MOX in H. contortus field isolate, as well as it may be used for official routine drug monitoring programs under the Ministry of Agriculture (MAPA) guidance.

Haemonchus contortus é uma das causas mais comuns e economicamente significativas de doença em produções de pequenos ruminantes em todo o mundo, e os programas de controle de parasitas nematoides - incluindo H. contortus - baseiam-se principalmente no uso de drogas anti-helmínticas. A consequência da utilização desses compostos, como sendo a única estratégia sanitária para evitar infecções por parasitas, tem sido a redução da eficácia de todos os produtos quimioterápicos, selecionando fortemente para resistência. O desenvolvimento generalizado da resistência anti-helmíntica e a dificuldade de seu diagnóstico precoce têm sido uma grande preocupação para o manejo sustentável de parasitas no campo. O objetivo desta pesquisa foi determinar e comparar o efeito da ivermectina (IVM) e da moxidectina (MOX) em um isolado de campo selecionado de H. contortus com um estado de resistência conhecido, utilizando o teste in vitro de migração de larvas em ágar (LMTA). Larvas de terceiro estágio de um isolado de H. contortus selecionado foram obtidas a partir de culturas de fezes de ovinos infectados experimentalmente e incubadas em onze concentrações diluídas crescentes de IVM e MOX (6, 12, 24, 48, 96, 192, 384, 768, 1536, 3072 e 6144µg/mL). As curvas sigmoides de dose-resposta foram obtidas utilizando o valor de R² >0,90 e a dose de concentração letal (CL50) para as drogas anti-helmínticas testadas, utilizando um modelo logístico de quatro parâmetros. O valor de CL50 para MOX foi significativamente menor do que para IVM (1,253µg/mL e 91,06µg/mL), identificando o isolado de H. contortus como consideravelmente menos suscetível à IVM em comparação à MOX. Além disso, o LMTA mostrou uma alta consistência (p<0,0001) e pode ser uma ferramenta útil de diagnóstico para monitorar o status de resistência de IVM e MOX em isolado de campo de H. contortus, assim como ser utilizado de forma oficial e em rotina para programas de monitoramento das drogas sob a demanda do Ministério da Agricultura (MAPA).

Evaluation of resistance in a selected field strain of Haemonchus contortus to ivermectin and moxidectin using the Larval Migration on Agar Test / Avaliação da resistência em um isolado de campo selecionado de Haemonchus contortus à ivermectina e moxidectina usando o Teste de Migração de Larvas em Ágar

Haemonchus contortus is one of the most common and economically significant causes of disease in small ruminants worldwide, and the control programs of parasitic nematodes - including H. contortus - rely mostly on the use of anthelmintic drugs. The consequence of the use of this, as the sole sanitary strategy to avoid parasite infections, was the reduction of the efficacy of all chemotherapeutic products with a heavy selection for resistance. The widespread of anthelmintic resistance and the difficulty of its early diagnosis has been a major concern for the sustainable parasite management on farms. The objective of this research was to determine and compare the ivermectin (IVM) and moxidectin (MOX) effect in a selected field strain of H. contortus with a known resistance status, using the in vitro larval migration on agar test (LMAT). Third stage larvae of the selected isolate were obtained from faecal cultures of experimentally infected sheep and incubated in eleven increasing diluted concentrations of IVM and MOX (6, 12, 24, 48, 96, 192, 384, 768, 1536, 3072 and 6144µg/mL). The dose-response sigmoidal curves were obtained using the R² value of >0.90 and the lethal concentration (LC50) dose for the tested anthelmintic drugs using a four-parameter logistic model. The LC50 value for MOX was significantly lower than IVM (1.253µg/mL and 91.06µg/mL), identifying the H. contortus isolate as considerably less susceptible to IVM compared to MOX. Furthermore, the LMAT showed a high consistency (p<0.0001) and provided to be a useful diagnostic tool for monitoring the resistance status of IVM and MOX in H. contortus field isolate, as well as it may be used for official routine drug monitoring programs under the Ministry of Agriculture (MAPA) guidance.(AU)

Haemonchus contortus é uma das causas mais comuns e economicamente significativas de doença em produções de pequenos ruminantes em todo o mundo, e os programas de controle de parasitas nematoides - incluindo H. contortus - baseiam-se principalmente no uso de drogas anti-helmínticas. A consequência da utilização desses compostos, como sendo a única estratégia sanitária para evitar infecções por parasitas, tem sido a redução da eficácia de todos os produtos quimioterápicos, selecionando fortemente para resistência. O desenvolvimento generalizado da resistência anti-helmíntica e a dificuldade de seu diagnóstico precoce têm sido uma grande preocupação para o manejo sustentável de parasitas no campo. O objetivo desta pesquisa foi determinar e comparar o efeito da ivermectina (IVM) e da moxidectina (MOX) em um isolado de campo selecionado de H. contortus com um estado de resistência conhecido, utilizando o teste in vitro de migração de larvas em ágar (LMTA). Larvas de terceiro estágio de um isolado de H. contortus selecionado foram obtidas a partir de culturas de fezes de ovinos infectados experimentalmente e incubadas em onze concentrações diluídas crescentes de IVM e MOX (6, 12, 24, 48, 96, 192, 384, 768, 1536, 3072 e 6144µg/mL). As curvas sigmoides de dose-resposta foram obtidas utilizando o valor de R² >0,90 e a dose de concentração letal (CL50) para as drogas anti-helmínticas testadas, utilizando um modelo logístico de quatro parâmetros. O valor de CL50 para MOX foi significativamente menor do que para IVM (1,253µg/mL e 91,06µg/mL), identificando o isolado de H. contortus como consideravelmente menos suscetível à IVM em comparação à MOX. Além disso, o LMTA mostrou uma alta consistência (p<0,0001) e pode ser uma ferramenta útil de diagnóstico para monitorar o status de resistência de IVM e MOX em isolado de campo de H. contortus, assim como ser utilizado de forma oficial e em rotina para programas de monitoramento das drogas sob a demanda do Ministério da Agricultura (MAPA).(AU)