Melatonin Administration to Pregnant Ewes for Coccidiosis Control in Their Offspring.

In livestock research, there has been a growing interest in the impact of melatonin on both health and disease conditions. The hypothesis of the present study was that melatonin treatment prenatally could support the immune competence and growth of experimentally infected lambs. This is the first study that aimed to investigate the impact of melatonin administration throughout pregnancy on immunity and oocyst excretion of pre-partum ewes and their offspring after experimental infection with Eimeria species. Thirty pregnant ewes were allocated into five equal groups, ΚΜ, ΚC, CM, CC, and NC, and gave birth to 47 lambs. Ewes of the KM and KC groups were orally challenged with a cocktail of Eimeria-sporulated oocysts (mainly consisting of Eimeria ovinoidalis), on day 120 of pregnancy, as well as all the lambs at the age of 5-9 days apart from those born from the NC group (environmental control). Fecal samples were collected from all ewes before infection and at parturition and from all lambs 14 times (S0-S13), before infection and during the following 8 weeks, for counting oocysts per gram of feces (OPG). Immunoglobulin (IgG) and cytokine (IL-1ß, IL-6, IL-10, IFN-γ) levels were determined in ewes' plasma collected before infection and at parturition, in lambs' plasma at 24 and 72 h after their birth, and in colostrum samples at parturition and 72 h later. Body weight of lambs was recorded five times from birth until the age of 60 days. Accordingly, the leucogram was evaluated in blood samples collected six times within the same period. On average, IgG concentration was higher (p < 0.05) in the blood of KM-ewes compared to KC and CC groups and in colostrum of KM-ewes compared to other groups (p < 0.001). KM-lambs had greater IgG titer and IFN-γ level than the other groups (p < 0.05). The IL-10/ IFN-γ ratio in KM-ewes was lower than the CC group (p = 0.06). Overall, the growth rate of lambs did not differ among groups (p > 0.05). Total oocysts' excretion in KM- and CM-lambs was reduced by 94.9% (p = 0.05) and 92.6% (p = 0.025), respectively, compared to KC-lambs, following the 3-week period after challenge, when E. ovinoidalis predominated in all groups. The dominant type of leucocytes was monocytes in all experimentally infected lambs, but not in NC-lambs, while overall lymphocytes were lower in KC-lambs than in NC-lambs (p < 0.05). Considering that almost all young indoor-reared lambs are exposed to coccidia species during their early life, melatonin treatment prenatally could suggest an alternative management tool in alleviating infection pressure.

Anguillicola (Anguillicoides) crassus: Morphometric characteristics and pathogenicity in eels (Anguilla anguilla) in Greece.

Anguillicola (Anguillicoides) crassus is a swimbladder nematode parasite of Anguilla anguilla (Linnaeus, 1758). The present study investigates the morphology and pathogenicity of A. crassus in European eel, as well as, the effects of different aquatic environments on the infection of A. crassus, in Greece. A total of 268 nematodes were collected from four estuarine systems in Greece. In all collected parasites, measurements carried out to define the morphological characteristics of parasites: parasite length and width, oesophagus max length and max width. Τhe mean parasite length was 23.50 mm (95% CI: 22.42-24.58) for females, 12.95 mm (95% CI: 12.25-13.64) for the males and 6.39 mm (95% CI: 5.27-7.50) for the juveniles. The mean parasite width was 1.99 mm (95% CI: 1.88-2.10) for females, 0.93 mm (95% CI: 0.86-1.00) for males and 0.51 mm (95% CI: 0.39-0.64) for juveniles. In total, the mean intensity was found 3.15 (95% CI: 2.53-3.78), while the highest mean intensity per eel was 8.00 (95% CI: 0.00-29.51) in Tholi Lagoon and the lowest was 2.20 (95% CI: 0.36-4.04) in Amvrakikos Gulf. Longitudinal and transverse histological sections of two adults A. crassus and two swimbladders were carried out. Multiple granules were observed, as a tissue reaction of the organism to swath around the 4th stage larvae (L4) that have entered the swimbladder. Molecular analysis was performed on three female adults A. crassus derived from eel specimens coming from the Vistonis estuarine system. The high reproductive capacity of the parasite reveals that A. anguilla has low effective defense mechanisms against the parasite. Also, the morphometric variations of the A. crassus seems to have a plastic feature which is being differently expressed when exposed to various environments.

Gastro-intestinal parasitism and control practices in dairy cattle in North-west Cameroon (Central Africa).

PURPOSE: The main objective of this study was to assess the gastrointestinal parasitism in relation with the worm control practices (WCP) in dairy cattle in areas of major importance for dairy/milk production of Cameroon. METHODS: Fresh fecal samples were randomly collected from 402 cattle from August to October 2016 in North West Cameroon, and analyze using the McMaster technique. Data on WCP were collected using a questionnaire. RESULTS: A total of 11 parasites species were identified: Trichostrongylus sp. (5.97%); Oesophagostomum sp. (5.47%); Haemonchus sp. (2.48%); Bonostomum sp. (1.74); Cooperia sp. (1.49%). Toxocara sp (0.24%); Ostertagia sp. (0.50%); Nematodirus sp. (0.74%); Trichuris sp. (0.50%); Moniezia sp. (0.50%); Eimeria sp. (0.50%). The mean individual parasite load for helminthes and protozoa ranged from 100 to 400 eggs per gram of feces (epg) and 333-400 oocysts per gram of feces (opg), respectively. Most farmers (95.12%) used conventional drugs (Albendazole, Doramectin, Fenbendazole, Ivermectin.) to deworm animals as compared with traditional phytotherapy (e.g., Carica papaya leaves) (4.88%). The mean epg/opg in animals treated with conventional drugs was significantly higher (p < 0.05) than in those receiving traditional treatment. Animals treated less than four times a year with conventional drugs had the highest (p < 0.05) mean epg/opg compared with animals treated four to six times a year. CONCLUSION: This study suggests that treating at least four times a year with conventional drugs, and at least two times a year with traditional approaches could help reducing GIP burden in dairy cattle in North West Cameroon. Since the majority of farmers use conventional drugs, it is imperative for them to appropriately use these conventional drugs for a better control of GIP on their farms.

Internal Parasites of Pigs and Worm Control Practices in Bamboutos, Western Highlands of Cameroon.

Internal parasites are limiting factors to successful, sustainable livestock production. Knowledge on how they are dealt with is important to prevent resistance to anthelmintics. The aim of this study was to describe the internal parasitism of indoor pigs in Bamboutos Division in Cameroon, as well as the attendant worm control practices. Thus, 324 pigs from 50 small scale farms were sampled for feces which were qualitatively and quantitatively examined for parasite eggs, cysts, or oocysts. Data on worm control practices were also collected. The overall prevalence was 74.7% (95 % Confidence Interval (CI): 69.6-79.3%) and the overall mean egg/oocyst per gram of feces (epg/opg) was 304.1±1218.0. The following parasites were found: Strongylid parasites (58.6%; epg= 105.0±134.7); Coccidia (26.9; opg=517.2± 1862.1); Strongyloides ransomi (25.9%; epg=61.9± 40.8); A. suum (3.7%. epg=50±0); Metastrongylus sp (0.9%; epg=50±0); Trichuris suis (0.9%; epg=50±0); and Macracanthorhynchus hirudinaceus (0.62%; epg=50±0). Single to septuple infestations occurred. The majority of farmers resorted to modern veterinary services (64%) and mostly used conventional drugs (88%). Internal parasitism was associated with the person in charge of animal health, the implementation of a prophylaxis program on the farm, and the annual deworming frequency. The implementation of a prophylaxis program significantly reduced the overall egg/oocyst load while high treatment frequency (more than thrice a year) did not, indicating that prophylaxis measures such as general hygiene must be reinforced in pig herds in the country, and the treatment frequency reduced as much as possible to prevent the selection of anthelmintic resistance.

An analysis of seroprevalence and risk factors for parasitic infections of economic importance in small ruminants in Greece.

A cross-sectional serological survey was carried out to screen the small ruminants of Thessaly, Greece, for infection with Haemonchus contortus, lungworms (i.e. Cystocaulus ocreatus, Muellerius capillaris, Protostrongylus rufescens and Neostrongylus linearis) as a group, and for Cysticercus tenuicolis, Linguatula serrata and Oestrus ovis. A second objective was to determine the risk factors related to herd characteristics, management practices, farmer status and bioclimatic variables associated with these parasitic infections. A total of 361 sheep and 179 goat serum samples were examined. The seroprevalences were 33.9%, 41.5%, 14.1%, 4.6% and 1.4% for H. contortus, lungworms, L. serrata, C. tenuicolis and O. ovis, respectively. The final logistic regression model showed that farm location and temperature were associated with H. contortus, lungworm and L. serrata infections. Anthelmintic treatment, class of anthelmintic and rotation of grazing were associated with H. contortus and lungworm infections, while grazing with other herds was associated with lungworm and L. serrata infections; rain was associated with H. contortus and L. serrata infections. Farm type and age of farmer were associated with H. contortus infections and elevation was associated with lungworm infection. The results may help to formulate appropriate control strategies in Greece and other areas with similar climatic conditions in order to channel limited resources to mitigate only those risk factors which are significant to protect the profitability of the livestock industry.

Global geographic distribution of Trichinella species and genotypes.

Maximum entropy ecological niche modeling was utilized to describe the global geographic distribution of Trichinella species and genotypes and to assess their invasive risk in new areas other than the ones currently known. Also, space-time scan statistic was utilized to identify global spatiotemporal clusters of infection. A database containing 3209 records for 12 species and genotypes identified at the International Trichinella Reference Center (ITRC) as well as climate, elevation, and land cover data extracted from various databases were used. Ecological niche modeling implemented in the Maxent program indicated new potential ranges for T. spiralis (T1), T. nativa (T2), T. britovi (T3), T. pseudospiralis (T4), T. murrelli (T5), T6, T. papuae (T10), and T. zimbabwensis (T11). The area under the curve values for the test data of the models ranged from 0.901 to 0.998, indicating that the models were very good to excellent. The most important bioclimatic factor in modeling the ranges for T. spiralis (T1), T. nativa (T2), T. britovi (T3), T6, and T. zimbabwensis (T11) was temperature, for T. pseudospiralis (T4) and T. papuae (T10) was precipitation, and for T. murrelli (T5) was land cover. T. spiralis (T1), T. britovi (T3), and T. pseudospiralis (T4) had the same primary land cover which was "Grass Crops". The primary land covers were "Conifer Boreal Forest" for T. nativa (T2), "Cool Fields and Woods" for T. murrelli (T5), "Upland Tundra" for T6, "Tropical Rainforest" for T. papuae (T10), and "Crops and Town" for T. zimbabwensis (T11). The scan statistic analyses revealed the presence of significant spatiotemporal clusters (p<0.05) for T. spiralis (T1), T. nativa (T2), T. britovi (T3), T. pseudospiralis (T4), T. murrelli (T5), T6, and T. nelsoni (T7). No significant clusters were found for T. papuae (T10) and T. zimbabwensis (T11).

Seroprevalence and risk factors associated with zoonotic parasitic infections in small ruminants in the Greek temperate environment.

A cross-sectional serological study was carried out to screen the sheep and goat population of Thessaly, Greece for evidence of infection with Toxoplasma, Toxocara, Leishmania, and Echinococcus and to determine the risk factors related to herd characteristics, herd management practices, farmer status, and the bioclimatic variables associated with these zoonotic parasitic infections. A total of 540 sheep and goat serum samples were examined. The seroprevalence of infection in all examined animals was 24.5% for Toxoplasma, 32% for Toxocara, 0% for Leishmania and 85.9% for Echinococcus. The final logistic regression model showed that the species of small ruminant, herd size, anthelmintic treatment, class of anthelmintic treatment, grazing with other herds, educational level of farmer, elevation of farm location, and generalized land cover were associated with Toxoplasma gondii infections, while the species of small ruminant, farm type, anthelmintic treatment, class of anthelmintic treatment, rotation of grazing, age of farmer, elevation of farm location, and generalized land cover were associated with Toxocara canis infections. Antibodies to T. gondii were detected in 102 (28.3%) of 360 sheep and in 30 (16.8%) of 179 goats. Animals in small flocks (150-300 animals) had an approximately 0.42-fold lower risk of having positive cases of T. gondii among animals compared with large flocks (>300 animals). Antibodies to T. canis were found in 155 (42.9%) of 361 sheep and 18 (10.1%) of 179 goats. The later finding constitutes the first report of seropositive goats to Toxocara. The risk of positivity for T. canis was 7.71-fold higher in sheep than in goats. Geographically, animals from plain areas had 2.9 and 2.01-fold higher risk of having positive cases of T. gondii and T. canis respectively. The significant bioclimatic variables (p<0.05) associated with the occurrence locations of T. gondii infection were related to higher temperature, lower precipitation, and lower elevation compared to the absence locations of T. gondii. The significant bioclimatic variables (p<0.05) associated with occurrence locations of T. canis infection were related to lower temperature and higher precipitation compared to absence locations of T. canis. These findings are useful to formulate appropriate control strategies for zoonotic parasites of sheep and goats in Greece and other areas with similar climatic conditions.

Geographic distribution modelling for ruminant liver flukes (Fasciola hepatica) in south-eastern Europe.

Maximum entropy ecological niche modelling was utilised to predict the geographic range for fluke genotypes and haplotypes in south-eastern Europe, using the Maxent program. The lowest (0.832) and the highest (0.947) area under the curve values were observed in the models for the haplotypes CtCmt1 and CtCmt2.2, respectively. Precipitation and temperature contribute equally to model building of the genotypes based on the 28S rDNA gene. In regard to the mtDNA gene region, precipitation is the most important factor in modelling the CtCmt1 haplotype range, while temperature appears to be the most important factor in modelling the CtCmt2.1 and CtCmt2.2 haplotype ranges. The highest level of probability for the geographic distribution of Fasciola hepatica genotypes and haplotypes covered the regions of southern Bulgaria and central and northern Greece which contain a high concentration of potential ruminant hosts.

Survey of gastrointestinal parasitic infections of beef cattle in regions under Mediterranean weather in Greece.

Gastrointestinal (GI) parasitic infections of beef cattle were investigated in 15 farms from 3 different regions of Greece, characterized by temperate Mediterranean climate, during two years. A total of 262 faecal samples were collected. Helminth eggs were detected in 42 (16%) samples. Strongyle-type eggs were found in 28 (10.7%) samples, Strongyloides spp. and Toxocara spp. eggs in 8 (3.1%) samples each, and Capillaria spp. and Moniezia spp. eggs in 1 (0.4%) sample each. Coccidian oocysts were found in 123 (46.9%) samples. A logistic regression model showed that the age of animals, the number of employees, and the region of animals were associated with coccidian infections. There was almost a four-fold increase in the risk of coccidian infections in calves less than 12 months old compared with animals that were more than 36 months old (p<0.05). The risk of coccidian infections was 0.07, 0.18 and 0.27-fold lower for farms with 1, 2 and 3 employees respectively compared to farms with 4 employees; farms with less than 4 employees had a protective effect (p<0.05 in all cases). Almost a four-fold increased risk of coccidian infections was shown for animals from Epirus compared with those from Thessaly (p<0.05). These findings are useful to formulate appropriate control strategies for coccidian parasites of beef cattle in Greece and other areas with similar climatic conditions.

Genetic diversity of equine piroplasms in Greece with a note on speciation within Theileria genotypes (T. equi and T. equi-like).

Equine piroplasms in Greece were studied using the reverse line blot hybridization (RLB) assay. Three genotypes consisting of two Theileria (T. equi and T. equi-like) and one Babesia (B. caballi-like) were identified. Of 787 samples tested, 371 (47.14%) hybridised to catchall probe (probe specifically designed to capture any piroplasm species present in a sample), 346 (43.96%) to T. equi probe, 364 (46.25%) to T. equi-like probe, 0 (0%) to B. caballi probe and 3 (0.38%) to B. caballi-like probe. Seven samples gave faint signals with the catchall probe only, indicating the presence of known or unknown piroplasm species, or a novel genotype or a known genotype occurring at a very low level of parasitemia. A partial sequence (509 bp) of the V4 region of the 18S rRNA gene of a T. equi-like isolate showed only 99% similarity with the reference T. equi-like isolates from Northern Spain from which the detecting probe used in the present study was designed but showed 100% similarity with the T. equi-like variants from Southern Spain. This indicated a noticeable degree of polymorphism within the population of T. equi-like. No unusual parasites previously reported in horses, such as B. canis canis and B. bovis were detected in this study. The values of the bioclimatic variables were very similar between the geographic locations for T. equi and T. equi-like genotypes, suggesting the two are not yet different species as hypothesized by some authors but are possibly undergoing a speciation process within Theileria genotypes. Both T. equi and T. equi-like were found in predominantly forest type land cover.

Geographic distribution modeling and spatial cluster analysis for equine piroplasms in Greece.

Maximum entropy ecological niche modeling and spatial scan statistic were utilized to predict the geographic range and to investigate clusters of infections for equine piroplasms in Greece, using the Maxent and SaTScan programs, respectively. The eastern half of the country represented the culminating area with high probabilities (p>0.75) of presence of equine piroplasms and encompassed most regions with high concentration of equid host populations. The most important environmental factor that contributed to the ecological niche modeling was land cover followed by temperature. Significant clusters (p<0.0001) were detected for Babesia caballi and Theileria equi infections in North and Central regions of Greece, respectively, which have significant equine populations. Maximum entropy ecological niche modeling and spatial scan statistic have proved to be useful tools for the surveillance of animal diseases.

Seroprevalence of equine piroplasms and host-related factors associated with infection in Greece.

Serum samples were collected from a total of 544 equids that included 524 horses, 13 mules, and 7 ponies from various regions of mainland Greece and were examined by competitive-inhibition ELISA (cELISA) to evaluate the level of exposure of Greek equids to Theileria (Babesia) equi and/or Babesia caballi, the causative agents of piroplasmosis. Association between seropositivity and host-related factors of species, gender, age, origin, activity and location were investigated. The overall seroprevalence was 11.6% (9.1-14.6%) with 95% confidence limit. The seroprevalence for T. equi and B. caballi was found to be 11% (8.6-14%) and 2.2% (1.2-3.9%), respectively. The animal-related factors significantly linked with seropositivity were the species, activities of farming, racing, recreation, and geographic location in Attica, Macedonia, Peloponnese and Thessaly region (p<0.05). The relative risks for the presence of T. equi, B. caballi and mixed infection in mules compared to horses was 8.39, 33.58 and 40.31, respectively. The infection level for T. equi, B. caballi and mixed infection were significantly higher in farm equids than in racing equids (p<0.05). Also, the rate of infection of T. equi was higher in farm equids than recreational equids (p<0.05). The relative risk of T. equi infection between farming equids and equids used only for recreation activity was 3.25-1, while the relative risk of B. caballi infection was 0.14-1 for racing animals relative to recreation animals. The region with the highest level of infection to both parasites was Thessaly (38.8% T. equi and 6.1% B. caballi), followed by Peloponnese (10.4% T. equi and 3.9% B. caballi), Attica region (8.3% T. equi and 0.6% B. caballi) and finally Macedonia the region with the lowest prevalence (6.6% T. equi and 4.4% B. caballi). A higher seroprevalence rate was found among local animals compared to imported equids, indicating that equine piroplasm infection is enzootic in Greece. T. equi seroprevalence was significantly different and higher among increasing age groups of equids, suggesting persistent infections or lower transmission levels whereby animals may need to be exposed longer before acquiring the infection. Competent tick vectors Rhipicephalus bursa and Rhipicephalus sanguineus for the transmission of equine piroplasmosis were recovered from horses and dogs, respectively.