Optimum timing for assessing phenotypic resistance against gastrointestinal nematodes in Pelibuey ewes.

The objective was to identify the optimal stage of production to evaluate the resistance of Pelibuey ewes against gastrointestinal nematodes (GIN). Faecal egg count (FEC) was used to classify the ewes as resistant, sensible or intermediate against GIN. Forty-seven ewes were mating during 30 d. The gestation was verified by ultrasonography, and the breeding date was used to calculate the productive stages. Faeces were taken weekly to determine the FEC. Blood samples were taken to determine the packed cell volume (PCV), the peripheral eosinophils count (PEC), plasma protein concentration (PP), and Immunoglobulin A (IgA) against Haemonchus contortus. The body condition score (BCS) was recorded at each visit. Six moments during the study (early, mid and late gestation; early, mid and late lactation) were considered. The ewes were classified according to FEC (mean FEC ± three standard errors). The higher FEC occurred during all lactation stages than during early and mid-gestation stages (P<0.05). PCV, PP, and BCS during early gestation stage were higher than shown during the lactation stages (P<0.01). The PEC and IgA were higher during all lactation stages than early and mid-gestation stages (P<0.05). Concerning the type of birth, double births showed higher FEC than single birth (P<0.01). The highest values of accuracy (100 %) and concordance (Youden's J = 1.0) were found during early lactation. Therefore, it is concluded that the optimal stage of production to evaluate phenotypic resistance against GIN infections in Pelibuey ewes was during the early lactation.

Identification of Somatic Proteins in Haemonchus Contortus Infective Larvae (L3) and Adults.

Haemonchus contortus is considered the most pathogenic nematode in sheep production systems based on grazing. Comparing infective larvae (L3) with adult parasites can lead to the identification of proteins that play an important role in parasite-host interactions. In this study, we report a list of H. contortus somatic proteins and made a comparative analysis of somatic proteins of L3 and adult worms. L3 and adult parasites were subjected to protein extraction and subsequently to peptide fractionation. Peptides were analysed by mass spectrometry and LC-MS/MS data analysis. Data analysis and search on SEQUEST and MASCOT against H. contortus from the WormBase ParaSite database resulted in the identification of 775 unique peptide sequences corresponding to 227 proteins at 1 % FDR. From these, 18 proteins were specific to L3 and 63 to adult parasites. The gene ontology (GO) enrichment analysis of the proteins specific to L3 and adult worms to gain insight into cellular components, molecular functions and biological processes that affect the parasite-host interaction showed some differences between the two parasite stages. The list of proteins found provides a database to identify target proteins that could be useful as biomarkers of the infection or in the generation of anthelmintic drugs that inhibit proteins essential for the establishment of the infection and the survival of adult parasites. They can also serve as new candidates for vaccine research.

Mean corpuscular haemoglobin concentration as haematological marker to detect changes in red blood cells in sheep infected with Haemonchus contortus.

Haemonchus contortus is a nematode parasite that causes anaemia and affects the health of sheep. The mean corpuscular haemoglobin concentration (MCHC) is an excellent indicator to detect anaemia that could help to characterize resistant or susceptible lambs to gastrointestinal nematodes. The aim of this study was to evaluate the predictive value of MCHC in detecting changes in red blood cells and their relation to anaemia in lambs re-infected with H. contortus. An analysis of information was performed using 24 Pelibuey lambs previously infected in grazing, dewormed and experimentally re-infected with H. contortus. At the first haematological sampling (admission) the lambs were classified based on MCHC quartiles (Q). Subsequently, the lambs were housed for 56 days. Blood samples were taken every seven days to determine the haematological parameters using an impedance haematological instrument. Confidence limits were constructed with the records of the lambs that recovered their haematological parameters. Each quartile was analysed as a treatment in a repeated measures design over time. To know the optimal combination of sensitivity and specificity of MCHC to detect anaemia a curve of receiver operating characteristic (ROC) curve and the cut-off values were evaluated. In quartile 4 (Q4), lambs showed the highest faecal egg count (FEC, 764 eggs/g of faeces), mean corpuscular haemoglobin (17.0 pg) and MCHC (54.6 g/dL). This group also presented the lowest RBC values (5.8 × 106/mL), haematocrit (HCT, 18.3%), total plasma protein (5.7 g/dL), and HGB (9.7 g/dL). The optimal point of MCHC with ROC curve was 42.4 (sensitivity 88.2% and specificity 86.5%); the area under the curve was 0.91 (CI 95%, 0.86-0.96). These results are related to the haematological effects caused by H. contortus in susceptible lambs. In conclusion, the highest FEC and lower HCT in Q4 are important elements of the haematological damage caused by H. contortus and could identify susceptible lambs.

Evaluation of anthelmintic drugs against egg development of rumen flukes recovered from cattle raised in the humid tropics of Mexico.

Paramphistomosis is a parasitic disease endemic in ruminants nearly worldwide. In the present study, an in vitro screening of the main anthelmintics used in Mexico was carried out to determine the mean lethal dose for rumen fluke eggs from cattle in a humid, warm region. Rumen flukes were obtained from cattle slaughtered in the states of Tabasco and Chiapas in Mexico. Eggs were collected using a 37-µm sieve and quantified. Then, an in vitro incubation study was performed: 100 eggs were placed into the wells of polystyrene microtiter plates. Anthelmintic products were tested on the eggs at concentrations ranging from 0.0015 to 3.0 mg/ml for rafoxanide, 0.0025 to 10.20 mg/ml for nitroxinil and 0.0015 to 3 mg/ml for closantel to determine the median lethal dose (LD50) and maximum lethal dose (LD99). A control group (water) was included in each plate. Three different species of rumen flukes (Calicophoron brothriophoron, Calicophoron clavula and Paramphistomum cervi) belonging to five isolates were identified. Nitroxinil had the highest efficacy against rumen fluke eggs, with an LD50 of 0.11 to 65 µg/ml, whereas rafoxanide showed the lowest efficacy with an LD50 ranging from 500 to 1713 µg/ml. Closantel showed high variability in the LD50 among the different analysed isolates (17 to 122 µg/ml). The evaluated flukicidal drugs presented differential efficacy against the development of rumen fluke eggs. The efficacy of the drugs will vary depending on the geographical area of origin of the animals.

Blood Feeding Status, Gonotrophic Cycle and Survivorship of Aedes (Stegomyia) aegypti (L.) (Diptera: Culicidae) Caught in Churches from Merida, Yucatan, Mexico.

Blood-feeding status, gonotrophic cycle, and survival rates of Aedes (Stegmyia) aegypti (L.) was investigated in catholic churches from Merida, Yucatan. Female Ae. aegypti were caught using backpack aspirator during 25 consecutive days in rainy (2015) and dry season (2016). Blood-feeding status was determined by external examination of the abdomen and classified as unfed, fed, and gravid. Daily changes in the parous-nulliparous ratio were recorded, and the gonotrophic cycle length was estimated by a time series analysis. Also, was observed the vitellogenesis to monitoring egg maturity. In total, 408 females Ae. aegypti were caught, and there was a significant difference in the number of females collected per season (Z = -6.729, P ≤ 0.05). A great number was caught in the rainy season (n = 329). In the dry season, 79 females were caught, which the fed females were twice greatest than the unfed. The length of gonotrophic cycle was estimated on the base of a high correlation coefficient value appearing every 4 days in rainy at 26.7 ± 1.22°C, and 3 days in dry season at 29.8 ± 1.47°C. The daily survival rate of the Ae. aegypti population was higher in both seasons, 0.94 and 0.93 for the rainy and dry season, respectively. The minimum time estimated for developing mature eggs after blood feeding was similar in both seasons (3.5 days in rainy versus 3.25 days in dry). The measurement of the vectorial capacity of Ae. aegypti in catholic churches could help to understand the dynamics of transmission of arboviruses in sites with high human aggregation.