Temporary Immunity of Blackbelly Lambs Reinfected with Trichostrongylus colubriformis

Background: Trichostrongylus colubriformis is one of the most prevalent gastrointestinal nematodes of sheep in warm climates. In this nematode species, anthelmintic resistance (AR) has been reported, which has motivated the search for alternative methods to control it. One practice is to increase the immune response level through the selection of animals with natural resistance, such as Blackbelly sheep. The aim of this study was to investigate the response of IgA and IgG in serum and saliva and the level of cells involved in the acquired resistance in Blackbelly lambs after single artificial reinfection with a resistant strain of Trichostrongylus colubriformis.Materials, Methods & Results: Sixteen weaned lambs grazed for one month in nematode-contaminated grasslands. All the lambs were treated with Albendazole and Levamisole; later, they were stabled for two months. After that, eight lambs were re-infected with 6000 larvae of T. colubriformis and other eight lambs remained naturally infected. In addition, eight nematode-free lambs raised in cages, served as negative control. Blood samples were taken fortnightly to determine packed cell volume (PCV, %), plasmatic protein (g dL-1), as well as absolute and differential leukocytes. The eggs per gram of feces (EPG) were recorded. An indirect ELISA against T. colubriformis Ag was performed to determine the IgG and IgA levels. All data were analyzed by repeated measures. The EPG in the re-infected group increased after 34 days (625 ± 287), while in the naturally infected group it remained close to 200 ± 158 throughout the study. The control lambs did not show any infection. The PCV (29.4 ± 3.4 %), basophils (44 ± 74), lymphocytes (8443 ± 2845) and monocytes (77 ± 91) counts were not affected by infection. The IgA OD against T. colubriformis increased significantly after 15 days post-infection (dpi) in the re-infected lambs (1.69 ± 0.5, 80% with regards to standard). With IgG was not possible to distinguish the effect of infection.Discussion: Highly persistent infection of T. colubriformis was observed for at least four months in the naturally infected group despite having been treated with anthelmintics. It is a significant problem because this species has a high prevalence in the region of study and shows anthelmintic resistance. For such reason, is important to evaluate the ability of the animal to develop an immune response. In other study, a natural reduction in the fecal egg count (FEC) was indicated at 100 dpi in the same species, but in the present work, the FEC could not be reduced until around 120 days when the naturally infected lambs were evaluated. The larger number of neutrophils and leukocytes in both the naturally infected and re-infected groups implies a sustained response for a long time. While the higher counts of eosinophils only in the re-infected group were produced as an immediate response to infection with T. colubriformis. Similarly, with a single infection of T. colubriformis, in Santa Ines sheep the infected lambs showed significantly higher levels of serum IgA than the control group. In this case, differences in IgA activity were observed between the re-infected lambs and the other groups at fifteen dpi, but the basal levels of IgA in the naturally pre-infected lambs were maintained at least five weeks after infection. In conclusion, an increase in the peripheral immune response in lambs re-infected with T. colubriformis resulted from the increase in IgA levels in serum and saliva, and increase in the number of peripheral eosinophils, total leukocytes and neutrophils. Re-infected lambs had a higher level of IgA than naturally infected lambs and control lambs, making possible to promote the immune response with a single re-infection with Trichostrongylus colubriformis

Fasciola hepatica and Rumen Flukes - In Vitro Evaluation of Main Commercial Anthelmintics

Background: Trematode infections are of great importance as they affect the health of many species of mammals as cattle, sheep and goat. Fasciola hepatica represents the main trematode zoonosis and risks to human and cattle and paramphistomosis is one emerging parasitic diseases of ruminants widely distributed in the world. The economic expenses are incurred by the use of ineffective anthelmintics for trematode control. Besides to faecal egg count reduction test (FECRT) to determine the anthelmintic efficacy, can be used in vitro assays, by this the aim of the study was to determine the lethal doses with hatching egg test of the main commercial anthelmintics used for the control of trematodes in cattle. Materials, Methods & Results: Liver and rumen were examined from cattle slaughtered in Tabasco, Chiapas and Campeche states from Mexico. F. hepatica eggs were recovered from gallbladder and rumen fluke eggs collected from adult parasites in saline solution. Subsequently, the hatching egg assays were performed placing 100 trematode eggs in distilled water in each one of 96 wells of polystyrene plates. After making the appropriate dilutions, several concentrations of commercial anthelmintics were evaluated, ranging from 0.04 to 80.63 mM for triclabendazole + 0.046 to 96.87 mM febendazole (TC+FBZ), from 0.04 to 91 mM for rafoxanide (RAFOX), from 0.02 to 43.74 mM for closantel (CLOS), from 0.036 to 76.18 mM for clorsulon + 0.002 to 3.31 mM ivermectin (CLORS+ IVM) and from 0.163 to 334.47 mM for nitroxynil (NITROX). A control group (water) was included in each plate. Lethal doses were obtained using the Probit procedure and analysis of the means with a one-way statistical design. Most drugs used against rumen fluke eggs presented a high LD50 and therefore were ineffective to cause egg mortality, such was the case of RAFOX that presented LD50 from 4,580 to 10,790 µg/mL (7 to 17 mM). CLOS presented the lowest LD50 (80 µg/mL or 0.12 mM) on rumen fluke eggs. TC+FBZ was found to be effective drug against the development of F. hepatica eggs in many samples. In the same way NITROX showed a low LD50 (37 to 63 µg/mL or 0.13 to 0.22 mM), but RAFOX presented a highest LD50 (1,450 µg/mL or 2.32 mM). Discussion: The present study focused on screen the ovicidal activity and determining in vitro lethal doses 50 of main commercial anthelmintics used to control F. hepatica and rumen fluke as rapid tests in a tropical region from Mexico. The FECRT is the main method to detect effectiveness of anthelmintic and other method is the coproantigen reduction test (CRT) by ELISA. Both tests require many infected animals depending the number of treatments and by this the egg hatch assay (EHA) represent a complementary diagnosis of effectiveness of anthelmintic products to compare between regions and even between farms, because few animals are required from the farm to collect trematode eggs, and it is possible to know the effectiveness against various anthelmintics at the same time. Efficacy studies on trematodes using egg hatching tests are scarce, although they have the advantage that they can be applied to both F. hepatica and rumen fluke. TC+FBZ was one of the most effective products in inhibiting the development of F. hepatica eggs. However, RAFOX showed low effectiveness against trematode eggs, with very high lethal doses. These results agree with a study that show low efficacy against the development of Paramphistomum cervi eggs and with the FECRT test reductions of 75% and 80.58% were obtained, in times from 7 to 84 days after treatment with RAFOX. NITROX and CLORS were drugs that had good efficacy on the development of F. hepatica eggs. A differential response between liver and rumen fluke was observed. The anthelmintics used against rumen fluke eggs show low ovicidal activity and in Fasciola hepatica TC+FBZ show the best activity. Keywords: trematodes, cattle, effectiveness, anthelmintics, ovicidal activity, egg hatching

Repetibilidade e variabilidade de medições de produção de metano e dióxido de carbono em bovinos alojados em câmaras de respiração de circuito aberto

Las pruebas de recuperación de gases son necesarias cuando se emplea la técnica de cámara de respiración para medir los gases de efecto invernadero exhalados por animales domésticos. Se obtuvo un conjunto de datos de 98 mediciones individuales de producción de metano y dióxidos de carbono de ganado alojado en dos cámaras de respiración para evaluar la recolección y repetibilidad de las mediciones realizadas. Se realizó un análisis de varianza para evaluar si existían diferencias estadísticamente significativas entre cámaras y entre animales. Los resultados mostraron las ocurrencias de variaciones en la produccion de metano entre las camaras.Estas variaciones pueden deberse muy probablemente a la fuga de aire de las cámaras oa las incertidumbres en los conductos de muestra de aire y las mediciones de flujo.Se requieren experimentos de recuperación de gases cuando la técnica de cámaras de respiración es empleada para la medición de gases de efecto invernadero exhalados por los animales domésticos. Un juego de 98 mediciones de metano y dióxidos de carbono producidos por bovinos alojados en dos cámaras de respiración fueron para estimar la escasez y repetibilidad de las mediciones realizadas. Se realizó un análisis de varianza de los datos para evaluar si existían diferencias estadísticamente significativas entre las cámaras y entre los animales. Los resultados mostraron las ocurrencias de variaciones en la medicion de metano entre las camaras.Estas variaciones pueden deberse probablemente a fugas de aire de las cámaras oa la incertidumbre en el ducto de prueba de aire o en la medición del flujo de aire.Experimentos de recuperação de gases são necessários quando a técnica da câmara de respiração é used to mid gases of efeito estufa exalados by animais domestics. Um conjunto de 98 medições de metano y dióxido de carbono producido por bovinos alojados em duas câmaras de respiração foi used to estimar a consumere e repetibilidad das medições used. Uma análise de variância dos dados foi realizado para avaliar se havia diferenças estatisticamente significativas entre as câmaras e entre os animais. Os resultados muestran una ocorrência de variações na medição de metano entre as câmaras. Essas variações são provavelmente devido a vazamentos de ar das câmaras ou incerteza no duto de amostragem de ar ou na medição do fluxo de ar

High Predatory Capacity of a Novel Arthrobotrys oligospora Variety on the Ovine Gastrointestinal Nematode Haemonchus contortus (Rhabditomorpha: Trichostrongylidae)

With the worldwide development of anthelmintic resistance, new alternative approaches for controlling gastrointestinal nematodes in sheep are urgently required. In this work, we identified and characterized native nematode-trapping fungi. We collected seven isolates of fungi with the capacity to form adhesive, three-dimensional networks as the main mechanism to capture, kill, and consume nematodes. The nematode-trapping fungi were classified into two groups; the first group includes the R2-13 strain, showing faster growth, abundant aerial hyphae, scarce conidia production, bigger conidia, and it formed a clade with Arthrobotrys oligospora sensu stricto. The second comprises the A6, A12, A13, R2-1, R2-6, and R2-14 strains, showing a growth adhering to the culture medium, forming little aerial hyphae, smaller conidia, and these formed a sister clade to A. oligospora. Except for the R2-6 strain, conidia production was induced by light. In all the strains, the predatory capacity against the sheep gastrointestinal nematode Haemonchus contortus was greater than 58% compared with the control group. The A6 and A13 strains were the most active against the infective H. contortus third instar (L3) larvae, with an average capture capacity of 91%. Altogether, our results support evidence for a novel A. oligospora variety with high nematode-trapping activity and promissory in helminthic controlThis research was funded by FONDEC-UAQ-2019 (Fondo para el desarrollo del Conocimiento—Universidad Autónoma de Querétaro, FNB202002) to E.U.E.N.S

Evaluación antihelmíntica de cuatro extractos de árboles forrajeros contra el nematodo Haemonchus contortus bajo condiciones in vitro

The objective was to evaluate the nematocidal effect of four hydroalcoholic extracts (HAE) of Brosimum alicastrum (HAE-Ba), Guazuma ulmifolia (HAE-Gu), Erythrina americana (HAE-Ea) and Leucaena leucocephala (HAE-Ll) against Haemonchus contortus. The tests of egg hatching inhibition (EHI) and larval (infective larvae) mortality were used. The treatments were HAEs at concentrations of 6.25-50 mg/mL for EHI and 25-100 mg/mL for larval mortality, ivermectin (5 mg/mL, positive control) and distilled water (negative control). Data were analyzed using an ANOVA and treatments with a concentration-dependent effect were subjected to a regression analysis to determine lethal concentrations (LC50 and LC90). In addition, a phytochemical analysis was performed on the extracts to identify the presence of the main secondary metabolites. The best ovicidal and larvicidal activity was observed in HAE-Gu with 96.78 % EHI at 6.25 mg/mL and 57.2 % larval mortality at 75 mg/mL. Followed by HAE-Ba showing 90 % EHI at 6.25 mg/mL and 58.0 % larval mortality at 75 mg/mL. The LC50 and LC90 of HAE-Gu on EHI were 2.7 and 4.4 mg/mL, respectively. While the LCs of this same extract on larvae were LC50= 64 and LC90= 125 mg/mL. The phytochemical analysis indicates that all extracts contain tannins, coumarins, flavonoids and terpenes. The fodder species G. ulmifolia and E. americana could be candidate plants for the control of H. contortus.El objetivo del presente estudio fue evaluar el efecto nematicida de cuatro extractos hidroalcohólicos (EHA) de Brosimum alicastrum (EHA-Ba), Guazuma ulmifolia (EHA-Gu), Erythrina americana (EHA-Ea) y Leucaena leucocephala (EHA-Ll) contra Haemonchus contortus. Se usaron las pruebas de inhibición de la eclosión de huevos (IEH) y mortalidad larval (larvas infectantes). Los tratamientos fueron los EHA´s a concentraciones de 6.25-50 mg/ml para IEH y de 25-100 mg/mlpara mortalidad larval, ivermectina (5 mg/ml, control positivo) y agua destilada (control negativo). Los datos se analizaron mediante un ANOVA y los tratamientos con efecto dependiente a la concentración se sometieron a un análisis de regresión para determinar las concentraciones letales (CL50 y CL90).  Además, se realizó un análisis fitoquímico a los extractos para identificar la presencia de los principales metabolitos secundarios.  La mejor actividad ovicida  y larvicida fue observada en  EHA-Gu con  un 96.78 % de IEH a 6.25 mg/ml y 57.2 % de mortalidad larval a 75 mg/ml. Seguido de EHA-Ba mostrando 90 % IEH a 6.25 mg/mly un 58.0 % de mortalidad larval a 75 mg/ml. Las CL50 y CL90 del EHA-Gu sobre la IEH fueron 2.7 y 4.4 mg/ml, respectivamente. Mientras que las CL´s de este mismo extracto sobre larvas fue de CL50=64 y CL90=125 mg/ml. El análisis fitoquímico indica que todos los extractos contienen taninos, cumarinas, flavonoides y terpenos. Las especies forrajeras G. ulmifolia y E. americana podrían ser plantas candidatas para el control de H. contortus

Efecto del hipoclorito de sodio y extracto de cítricos en la reducción de la infestación con nematodos gastrointestinales resistentes a antihelmínticos en ovinos de pelo

The study included two experiments. The objective of experiment one was to learn about the in vitro effect of different sodium hypochlorite concentrations and a commercially-available citric extract on the survival of infestant larvae of gastrointestinal nematodes by observation at different time points. With >0.05% sodium hypochlorite concentrations, larvae lost their sheaths in two minutes, while at 1.3% concentration larvae died within one hour. Larval mortality also occurred as late as 12 h after exposure to the concentrations tested of citric extract.El estudio se realizó en dos experimentos. En el primero el objetivo fue conocer el efecto in vitro de diferentes concentraciones de hipoclorito de sodio y de un extracto de cítricos comercial sobre la sobrevivencia de larvas infestantes de nematodos gastrointestinales, mediante observación a diferentes tiempos. A concentraciones mayores al 0.05 % de hipoclorito de sodio, las larvas se desenvainaron en dos minutos y con concentraciones de 1.3 % las larvas murieron en una hora. Con el extracto de cítricos se observó mortalidad larvaria hasta las 12 h en todas las concentraciones utilizadas

Comparison of two phenotypical methods to segregate resistant and susceptible lambs to parasitic nematodes

The objective of this study was to compare two segregation methods to select resistant and susceptible female Pelibuey lambs infected naturally with gastrointestinal nematodes (GINs) in relation to their haematological and immunological response. For 6 months, faeces and blood samples were taken fortnightly from 40 grazing 5-month-old female lambs. The lambs were classified according to two methods using faecal egg count (FEC) as a phenotypical trait. In the first (reference) method (M3SE, n = 22), resistant (RES) lambs had FEC lower than the mean – 3 standard errors, the susceptible (SUS) lambs levels higher than + 3 standard errors(n = 10) and the intermediate (INT) lambs (n = 8) were categorised by having FECs between the two values. The second method (QUM) divided the population, using quartiles, into resistant (RES; 25%), intermediate (INT; 50%), and susceptible (SUS; 25%) lambs. The agreement between both methods was estimated using the Kappa index. The packed cell volume (PCV), total plasma protein (TPP) and peripheral eosinophils (EOS) were determined for each group. Serum was used to evaluate the IgA levels. PCV and TPP values were higher (P<0.01) in the RES lambs (31.5 ± 3.4 and 6.16 ± 0.5 g/dL by QUM, respectively, and 31.5 ± 3.9 and 6.24 ± 0.49 g/dL by M3SE, respectively) than the SUS lambs (28.1 ± 4.7 and 5.94 ± 0.5 g/d, respectively, by both methods). The EOS and IgA values increased with age. M3SE and QUM were in moderate agreement (Kappa = 0.43). We concluded that the two segregation methods allowed for the identification of the same female SUS lambs, but a greater number of animals were categorised phenotypically as resistant using the M3SE method. PCV and TPP can help to identify phenotypically resistant animals

Evaluación de parámetros productivos y reproductivos en un hato de doble propósito en Tabasco, México

El objetivo del estudio fue evaluar el comportamiento productivo y reproductivo de bovinos Bos taurus (BT) x Bos indicus (BI) en un sistema de doble propósito, bajo las condiciones tropicales del estado de Tabasco. Los registros se obtuvieron de la unidad de producción, UACH-URUSSE, ubicada en Teapa, Tabasco. Los años evaluados fueron 1994 a 2011, presentando información de 492 partos, y 240 lactancias completas de 130 vacas Cebú o sus cruzas (BT x BI). Los genotipos presentes en este estudio fueron las cruzas de Cebú (C) x Holstein (H): 7/8C1/8H, 3/4C1/4H, 5/8C3/8H, 1/2C1/2H, 5/8h3/8C, 11/16H5/16C y 3/4H1/4C; estos se agruparon en tres categorías 0-25%, 37.5-50% y 62.5-75% de acuerdo a la proporción de genes BT. Las variables incluyeron peso al nacimiento (PN), intervalo entre partos (IEP), producción de leche por lactancia (PLL) y duración de lactancia (DL). Los resultados indicaron que los genotipos con mayor proporción de genes BT influyó favorablemente sobre PLL (P0.05). El genotipo 62.5-75% presentó la mejor PLL (1,262.80 ± 34.80 kg) y la DL más prolongada (260.34 ± 11.20 días); comparada con los genotipos 0-25% y 37.5-50%, PLL fue 8.09 y 6.56 % mayor y con DL fue 41.72 y 33.68 días mayor, respectivamente. En conclusión, el genotipo con 62.5-75% de genes BT presentó el mejor comportamiento productivo; sin embargo, el comportamiento reproductivo fue similar en los genotipos estudiados

Inuencia materna en el crecimiento predestete y características de la canal de corderos de pelo/Maternal inuence on preweaning growth and carcass characteristics of hair lambs

El objetivo del presente estudio fue evaluar la inuencia del tipo de nacimiento (TN) de los corderos y el número de parto (NP) de las ovejas sobre el crecimiento predestete, rendimiento de la canal (RC), composición corporal de los corderos y el comportamiento productivo de las ovejas. Se estudiaron 25 ovejas Blackbelly x Pelibuey y sus corderos, bajo un diseño factorial, los factores fueron TN (sencillo y múltiple) y NP (primípara y multípara). Los corderos se sacricaron a los 56 d de edad. Las variables evaluadas fueron: peso vivo al nacimiento (PN) y al destete (PD), ganancia diaria de peso (GDP), peso de la canal y RC, peso y rendimiento de los cortes primarios. En las ovejas, se evaluó el peso al parto y al nalizar la lactancia, cambio de peso (CP), peso de la camada al nacimiento (PCN) y al destete (PCD). El TN afectó (p < 0.05) la GDP, PD, peso y RC de los corderos. En las ovejas, el tipo de parto (TP) y NP inuyeron (p < 0.05) sobre el CP. El TP afectó (p < 0.05) el PCN y PCD. El TN afectó el crecimiento predestete, RC y composición corporal de los corderos, siendo los de nacimiento sencillo los que tuvieron mayor GDP, peso y RC. Mientras que las ovejas multíparas y ovejas con parto múltiple experimentaron el mayor CP negativo. En tanto que las ovejas con parto múltiple mostraron la mayor eciencia productiva