Molecular identification of Chrysomya albiceps (Diptera: Calliphoridae) in a case of primary cutaneous myiasis in sheep at southern Brazil.

Sheep farming has been growing in Brazil, driven by an expanding consumer market due to greater acceptance of its meat and derivatives. There are several factors that limit sheep production, and one of them is infestation by ectoparasites, which cause stress in animals, weight loss, poor development, low productivity, low quality wool and reduced fertility. Chrysomya albiceps is a species of blowfly belonging to the Calliphoridae family that occurs in neotropical regions, where it causes secondary myiasis. We identified here a rare case of cutaneous myiasis with the presence of tissue lesions caused by C. albiceps in sheep in southern Brazil. We highlight the need to carry out more in-depth studies regarding the biology of these insects, with the aim of proving this atypical behavior for Brazil.

Treatment and management of coenurosis by Taenia multiceps: field data from outbreaks in endemic regions and literature review.

BACKGROUND: Taenia multiceps coenurosis is endemic in sheep from various regions worldwide. Dogs, the key hosts, shed T. multiceps eggs in their feces contaminating the pasture, and lambs are mostly infected during their first turnout into pastures. The disease is manifested in two forms: acute (due to the migrating oncospheres in the CNS) or chronic (due to the developing coenuri in the brain or spinal cord). Both forms are frequently accompanied by neurological symptoms. METHODS: Field trials conducted in an endemic region (Sardinia, Italy) to treat replacement lambs in six sheep flocks infected with acute coenurosis are summarized in this article. The article also reviews earlier reports on various approaches developed to treat and immunize sheep against coenurosis. RESULTS: Accurate detection of the time in which lambs become infected is crucial in deciding which treatment approach should be used. Acute disease can be successfully treated via chemotherapy. Results of field trials conducted in Sardinia revealed the efficacy of three (1-week apart) oxfendazole doses (14.15 mg/kg) in protecting apparently healthy lambs in the infected flocks from developing neurological symptoms. A single praziquantel dose (18.75 mg/kg) worked well for the same purpose and was also found significant in treating 5 of 16 clinically ill lambs in one flock. Earlier reports documented high rates of recovery (up to 100%) in clinically diseased lambs that received much higher doses (50-100 mg/kg) of praziquantel. However, chemotherapy is not preferred in chronic coenurosis since it can lead to rupture of the coenuri, giving rise to serious inflammation in the CNS. Surgical intervention is highly recommended in this case, and the pooled success rates for surgery in chronic-infected cases was estimated at 82.1% (95% CI 73.1-91.0%). However, various trials have been conducted to immunize sheep against T. multiceps coenurosis, and the 18k (Tm18) family of oncosphere antigens was found promising as a vaccine candidate. CONCLUSIONS: In acute coenurosis, selection of the proper anthelmintic should be done after consulting the owner for several reasons: (1) costs of the used anthelmintic: treating a small flock of 100 sheep costs around 1170 and 660 € for praziquantel and oxfendazole, respectively; (2) withdrawal time of the used anthelmintic: No time is required before consuming meat and milk from praziquantel-treated sheep, whereas meat and milk from oxfendazole-treated sheep should not be consumed for 44 and 9 days, respectively, causing additional costs for the farmers. Since no commercial vaccines have yet been developed against T. multiceps coenurosis in sheep, preventive measures remain the cornerstone of controlling this serious disease.

A pilot study of the in vitro efficacy of different concentrations of Duddingtonia flagrans for the control of gastrointestinal nematodes of sheep.

Duddingtonia flagrans is a nematode trapping fungus used for the control of gastrointestinal nematodes in livestock. The quantity of chlamydospores of D. flagrans required for the reduction of third-stage larvae (L3) of sheep gastrointestinal nematodes (GIN) is largely unknown, and a matter of discussion. The aim of this experiment was to determine in vitro the nematophagous activity of four different concentrations of D. flagrans (1000, 3000, 6250, or 11000 chlamydospores/ml) in the presence of varying numbers of GIN third-stage larvae (L3) (500, 1000, 1500). Additionally, the study sought to evaluate the efficacy of this fungus on Haemonchus contortus, Teladorsagia circumcincta, Trichostrongylus colubriformis and Chabertia ovina. The results showed that as fungal concentrations increased, so did the larval reduction of third-stage infective larvae in each test. L3s number was not a determining factor in the efficacy against GIN. The comparison between various concentrations of chlamydospores revealed significant differences, particularly between 1000 and 11000 chlamydospores (P≤0.05). Regarding the larval reduction of the GIN species considered, D. flagrans demonstrated the same effectiveness across all species tested. The results of the current study confirm the efficacy and underscore the importance of D. flagrans as an alternative for controlling of GIN.

High genetic diversity of Anaplasma ovis in sheep from Bosnia and Herzegovina.

BACKGROUND: Ovine anaplasmosis (sensu stricto) is a rickettsial blood disease caused by the tick-borne species Anaplasma ovis. The disease is characterized by mild anemia, fever, and icterus. A more severe clinical presentation is possible in non-endemic areas. There is no existing data on the presence of Anaplasma ovis in Bosnia and Herzegovina. However, given the country's location within the Mediterranean Basin and the recent molecular detection of Babesia ovis, it is plausible that sheep in the region could naturally be infected with this tick-borne pathogen. METHODS AND RESULTS: Blood samples from 81 sheep in the Podrinje and Herzegovina areas were examined by PCR. PCR positivity was found in 38 (46.9%) cases indicating a high number of infected sheep. Mixed infections with Babesia ovis and A.ovis were observed in 63.3% of cases. A higher number of positive sheep was recorded in the area of Herzegovina. Phylogenetic analysis of the gltA, groEL, and msp4 genes of A. ovis revealed numerous genotypes and significant genetic variability. This diversity was not related to geographic origin, tick-borne infection status, or sheep breeding practices in Podrinje and Herzegovina. CONCLUSIONS: The data obtained in this study suggest that the emergence of new genotypes and the high genetic variability of A. ovis are driven by specific local and micro-environmental factors.

Role of Rhipicephalus bursa larvae in transstadial transmission and endemicity of Babesia ovis in chronically infected sheep.

Babesia ovis, transmitted by Rhipicephalus bursa ticks, is the causative agent of ovine babesiosis, a disease characterized by fever, anemia, hemoglobinuria, and high mortality in sheep. This study investigates whether sheep that survived babesiosis without treatment can serve as a source of infection for B. ovis-free host-seeking R. bursa larvae in a later season. Three donor sheep were experimentally infected with B. ovis, and after six months, persistence of B. ovis was assessed through blood and tick transmission experiments. Blood from donor sheep was intravenously injected into three recipient sheep, while donor sheep were also infested with B. ovis-free R. bursa larvae. Engorged nymphs molted to adults, and new recipient sheep were infested with these ticks. All recipient sheep were monitored for B. ovis for 100 days using microscopic, serological, and molecular approaches. The presence of B. ovis was confirmed in the recipient sheep that received blood, leading to clinical infection in two. However, no B. ovis was detected in recipient sheep infested with ticks. These results suggest that sheep recovering from B. ovis infection do not serve as a source of infection for R. bursa larvae in subsequent seasons.

Prevalence and subtyping of Blastocystis sp. in ruminants in Southwestern, Iran.

Blastocystis is the most common gastrointestinal protozoan parasite of humans and many vertebrates. This study was carried out to investigate the prevalence and determination subtype (ST) of Blastocystis in domestic ruminants of Shahrekord County, southwestern Iran. In this descriptive cross-sectional study, 330 ruminant fecal samples (107 cows, 115 sheep, and 108 goats) were evaluated by parasitological methods (direct wet mount microscopic examination and formalin-ether concentration), Giemsa staining, In vitro xenic culture (The modified Dobell and Laidlaw culture method), polymerase chain reaction, and sequencing from 2018 to 2019, then data were analyzed using SPSS software version 21. The overall Blastocystis positive in ruminants was 14.2% and the frequency of Blastocystis sp. in cattle, sheep, and goats were 0.93%, 17.4%, and 24.1% respectively. Molecular diagnosis techniques revealed that ruminants were infected with four STs (genotypes) of Blastocystis including ST5(21.3%), ST7(2.1%), ST10(17.1%) and ST14(57.4%). Also, the STs identified in cows were ST10, and the observed STs in sheep were ST5 (40%), ST7 (3%), ST10 (5%), ST14 (45%), and one unknown subspecies. Goats were infected by ST5 (7.7%), ST10 (23.1%), and ST14 (69.2%). In this study, ST14 was identified as the most common subtype of Blastocystis sp. that was not common between humans and livestock, meanwhile, ST5 and ST7 are common between humans and animals accounted 21.3% and 2.1% of the positive cases, respectively, and reinforces the hypothesis that ruminants are reservoirs of blastocystosis in humans.

Metagenome reveals the midgut microbial community of Haemaphysalis qinghaiensis ticks collected from yaks and Tibetan sheep.

BACKGROUND: Haemaphysalis qinghaiensis is a tick species distributed only in China. Due to its ability to transmit a variety of pathogens, including species of the genera Anaplasma, Rickettsia, Babesia, and Theileria, it seriously endangers livestock husbandry. However, the microbial community of the midgut of H. qinghaiensis females collected from yaks and Tibetan sheep has not yet been characterized using metagenomic sequencing technology. METHODS: Haemaphysalis qinghaiensis were collected from the skins of yaks and Tibetan sheep in Gansu Province, China. Genomic DNA was extracted from the midguts and midgut contents of fully engorged H. qinghaiensis females collected from the two hosts. Metagenomic sequencing technology was used to analyze the microbial community of the two groups. RESULTS: Fifty-seven phyla, 483 genera, and 755 species were identified in the two groups of samples. The ticks from the two hosts harbored common and unique microorganisms. At the phylum level, the dominant common phyla were Proteobacteria, Firmicutes, and Mucoromycota. At the genus level, the dominant common genera were Anaplasma, Ehrlichia, and Pseudomonas. At the species level, bacteria including Anaplasma phagocytophilum, Ehrlichia minasensis, and Pseudomonas aeruginosa along with eukaryotes such as Synchytrium endobioticum and Rhizophagus irregularis, and viruses such as the orf virus, Alphadintovirus mayetiola, and Parasteatoda house spider adintovirus were detected in both groups. In addition, the midgut of H. qinghaiensis collected from yaks had unique microbial taxa including two phyla, eight genera, and 23 species. Unique microorganisms in the midgut of H. qinghaiensis collected from Tibetan sheep included two phyla, 14 genera, and 32 species. Kyoto Encyclopedia of Genes and Genomes enrichment analysis revealed that the functional genes of the microbiome of H. qinghaiensis were annotated to six pathways, and the metabolic pathways included 11 metabolic processes, in which the genes involved in carbohydrate metabolism were the most abundant, followed by the genes involved in lipid metabolism. CONCLUSIONS: These findings indicate that most of the microbial species in the collected H. qinghaiensis ticks were the same in both hosts, but there were also slight differences. The analytical data from this study have enhanced our understanding of the midgut microbial composition of H. qinghaiensis collected from different hosts. The database of H. qinghaiensis microbe constructed from this study will lay the foundation for predicting tick-borne diseases. Furthermore, a comprehensive understanding of tick microbiomes will be useful for understanding vector competency and interactions with ticks and midgut microorganisms.

Optimizing sheep B-cell epitopes in Echinococcus granulosus recombinant antigen P29 for vaccine development.

Background: Echinococcus granulosus is a widespread zoonotic parasitic disease, significantly impacting human health and livestock development; however, no vaccine is currently available for humans. Our preliminary studies indicate that recombinant antigen P29 (rEg.P29) is a promising candidate for vaccine. Methods: Sheep were immunized with rEg.P29, and venous blood was collected at various time points. Serum was isolated, and the presence of specific antibodies was detected using ELISA. We designed and synthesized a total of 45 B cell monopeptides covering rEg.P29 using the overlap method. ELISA was employed to assess the serum antibodies of the immunized sheep for recognition of these overlapping peptides, leading to the preliminary identification of B cell epitopes. Utilizing these identified epitopes, new single peptides were designed, synthesized, and used to optimize and confirm B-cell epitopes. Results: rEg.P29 effectively induces a sustained antibody response in sheep, particularly characterized by high and stable levels of IgG. Eight B-cell epitopes of were identified, which were mainly distributed in three regions of rEg.P29. Finally, three B cell epitopes were identified and optimized: rEg.P2971-90, rEg.P29151-175, and rEg.P29211-235. These optimized epitopes were well recognized by antibodies in sheep and mice, and the efficacy of these three epitopes significantly increased when they were linked in tandem. Conclusion: Three B-cell epitopes were identified and optimized, and the efficacy of these epitopes was significantly enhanced by tandem connection, which indicated the feasibility of tandem peptide vaccine research. This laid a solid foundation for the development of epitope peptide vaccine for Echinococcus granulosus.

Prevalence of Fasciola hepatica infection in slaughtered sheep from Northwest Tunisia and its risk factors: Association with gastrointestinal helminths infection and anaemia.

We investigated herein the prevalence of Fasciola hepatica infection in sheep at Sejnane slaughterhouse, governorate of Bizerte, Northwest of Tunisia, using three different diagnostic techniques (liver dissection, bile examination, and coprology). Faeces, liver, gall bladder as well as blood samples were collected from 603 slaughtered sheep in two seasons: winter and summer. Faecal egg counts of F. hepatica were estimated using sedimentation technique. Livers were examined for the presence of flukes, and bile collected from gall bladder was examined by sedimentation technique for the presence of F. hepatica eggs. Faecal egg counts of gastrointestinal helminths were estimated using flotation followed by the McMaster technique. Blood samples were used to estimate blood cell count (RBC) (×106/mL), haemoglobin (Hb) (g/dL), and haematocrit (Ht) (%) levels. A total of 1714 F. hepatica flukes were collected from 68 infected livers, the number of flukes per sheep ranged between naught and 195. Bile examination (16.78% ± 1.83; 51/310) showed the higher infection prevalence, followed by liver dissection (11.28% ± 1.17; 68/603) and coprology (9.12% ± 1.08; 55/603) (p = 0.015). Infection prevalences were significantly higher in young sheep aged of less than 1 year (8.13% ± 1.22; 49/498), in cross-bred sheep (10.61% ± 1.39%; 64/478), and in summer (7.13% ± 1.82; 43/293) (p < 0.05). There was no significant difference in infection prevalence by gastrointestinal helminths in F. hepatica-infected and F. hepatica-non-infected animals (p > 0.05). The overall prevalence of F. hepatica-infected anaemic sheep was higher (22.73% ± 4.47; 20/88) than F. hepatica-non-infected anaemic sheep (p < 0.05). Fasciola hepatica infection is frequent in sheep from Sejnane representing hence an important constraint for the development of the sheep industry in this region. Therefore, it is necessary to establish and implement a specific control programme to reduce fasciolosis infection risks including animal owners' education.

Expression of Toll-like receptors in Haemonchus Contortus resistant sheep: An innate immune parameter for host defense against gastrointestinal nematode infection.

Innate immune parameters, a first line of defense against invading pathogens like bacteria, parasites, fungi, etc, play a significant role in the prevention and elimination of aetiological agents primarily by recognition of invading pathogen-specific molecules by different pattern recognition receptors. Toll-like receptors (TLRs), a type-I transmembrane glycoprotein, cause innate immune responses mainly by produing inflammatory cytokines, chemokines and interferons. The objective of present study was to determine the role of TLRs in parasite resistance in Malpura sheep. In the current study, transcript variation of TLRs and its downstream signalling molecules namely MyD88, TRIF, IRF-3, TRAF, TGF-ß, NFκB, and CD14 were ascertained by real-time PCR in Haemonchus contortus resistant (R) and susceptible (S) Malpura sheep. Results have shown significantly (P<0.05) up-regulated expression of TLR-2, TLR-4, TLR-5, TLR-8 and TLR-10 in July however down-regulated patterns were observed in August and September in R-line sheep compared to S-line sheep. This indicates that at more or less equal parasite load, the TLR genes in R sheep produce more transcripts, but after parasite loads have increased hugely in the S line, they easily surpass the levels seen in the S line. Result suggests that transcriptional activity of the TLR genes was related to parasite load and there were differences between the lines at different infection intensities. Three-point transcript expression observation of the signalling molecules namely TRIF, IRF-3, TRAF, a similar pattern was observed in R sheep compared with S sheep.

Investigation of Toxoplasma gondii Infection in Yunnan Semi-fine Wool Sheep (Ovis aries) and wild Rodents in Yunnan, China.

BACKGROUND: Toxoplasma gondii, a globally distributed zoonotic obligate intracellular parasite, infects a wide array of mammals, including humans, sheep, and birds. As a unique sheep breed in southwestern China, Yunnan semi-fine wool sheep occupies an important position in animal husbandry in Zhaotong due to its strong adaptability, high reproductive rate, and excellent wool quality. Lambs infected with T. gondii are prone to neurological symptoms and growth retardation, while T. gondii infection in ewes can cause abortions, stillbirths, and deformities, thus affecting sheep reproduction and sheep product quality. Meanwhile, mutton and dairy products contaminated with T. gondii can become potential sources of human infection, potentially threatening public health and safety. METHOD: To understand the T. gondii infection in semi-fine wool sheep in Zhaotong, Yunnan Province, 586 blood samples were collected and subjected to indirect hemagglutination assay (IHA) for T. gondii antibodies, and the infection-related factors were analyzed through cross-sectional analysis. In the meantime, nested PCR was conducted on a total of 217 samples collected from 31 rodents caught in and around the sheep breeding ground to test the T. gondii B1 gene in rodent tissues. RESULTS: A total of 94 sera tested positive for T. gondii antibodies, with a total positive rate of 16.04% (94/586) (95% CI: 14.77-20.89). Cross-sectional statistical analysis on factors related to semi-fine wool sheep infection rate, including sampling season, sex, age, and weight, suggested that age (< 6 months: 23.81%; 6-12 months: 11.74%; > 12 months: 15.83%) was a significant factor explaining the infection rate differences (P = 0.003 < 0.05, χ2 = 11.62, df = 2). Thus, age was considered a key risk factor for T. gondii infection in this study (odds ratio, OR = 2.35, 95% CI: 1.42-3.87). Nested PCR analysis on 217 (heart, liver, spleen, lung, kidney, brain, and muscle) tissues from the 31 rodents indicated that 11 tested positive. The total infection rate of rodents in and around the breeding ground was 35.48% (11/31), and 14 samples tested positive, with a positive infection rate of 6.45% (14/217). CONCLUSION: The T. gondii infection rates of semi-fine wool sheep and rodents from their breeding environment in Zhaotong, Yunnan Province, were high, necessitating enhanced prevention, control, and treatment measures to ensure the healthy breeding of semi-fine wool sheep and veterinary public health and safety.

First Molecular and Phylogenetic Identification of Toxoplasma Gondii in Sheep Liver Intended for Human Consumption in Northern Tunisia.

The aim of this study was to estimate the molecular infection prevalence of Toxoplasma gondii in sheep liver tissues destined for human consumption. A total number of 224 liver tissues were collected from slaughtered sheep in Sejnane slaughterhouse (Northwest Tunisia). PCR was used to detect T. gondii DNA in liver tissues followed by phylogenetic analysis of amplicons. The phylogenetic tree was then constructed to compare the partial sequences of the ITS1 gene with GenBank sequences.The overall molecular prevalence of T. gondii in sheep livers was 25% (56/224). The highest molecular prevalence of T. gondii was recorded in sheep aged of less than one year old (27.3%; 52/190). Infection prevalence was significantly higher in Noire de Thibar breed (33%; 17/51) compared to other breeds (p = 0.023). There were no differences depicted according to sheep's gender. The T. gondii sequences obtained in the present study (GenBank accession numbers: OR509829 and OR509830) were 98.40-100% homologous to T. gondii sequences published in the GenBank. These results highlight a high level of T. gondii contamination of tissues destined for human consumption. Further studies are needed to improve our knowledge on different genotypes of T. gondii that infect Tunisian sheep population.

The first molecular detection of benzimidazole resistance in Haemonchus contortus from sheep in Hejing and Minfeng counties of Southern Xinjiang.

To understand the benzimidazole (BZ) resistance of Haemonchus contortus in Southern Xinjiang, three single nucleotide polymorphisms (SNPs) designated as F167Y, E198A, and F200Y, in the isotype-1 ß-tubulin gene which are associated with BZ resistance, were investigated for H. contortus populations from sheep in Hejing and Minfeng counties of Southern Xinjiang. In brief, a total of 190 H. contortus adults were collected from 52 out of 70 slaughtered sheep in city abattoirs across two regions in Southern Xinjiang. The species identity of each adult worm was confirmed by PCR amplification of ITS-2 using H. contortus-specific primers targeting the ITS-2. The samples were then investigated for BZ-related SNPs at locus 167, 198, and 200, by PCR-sequencing of the isotype-1 ß-tubulin gene. The results showed that only E198A and F200Y mutations were detected in the investigated H. contortus populations. The E198A mutation (homozygous and heterozygote resistant: found in 40% and 30% of sequenced samples from Minfeng and Hejing counties, respectively) was predominant compared with the F200Y mutation (homozygous and heterozygote resistant: found in 14% and 13.3% of sequenced samples from Minfeng and Hejing counties, respectively). The results indicate a high prevalence of BZ resistance in H. contortus populations from certain areas of Southern Xinjiang. Our findings provide valuable information for the prevention and control of H. contortus in areas with similar conditions.

Molecular prevalence, associated risk factors and phylogenetic evaluation of Theileria lestoquardi in the blood samples of small ruminants.

Raising small ruminants is the main source of income for farmers in Pakistan especially in rural areas of Dera Ghazi Khan in Punjab. Despite having large sheep population, the prevalence of intra-erythrocytic protozoa, Theileria (T.) lestoquardi, has never been reported from this area. This study was conducted to fill this knowledge gap and 333 blood samples of apparently healthy small ruminants (168 sheep and 165 goats) along with their epidemiological data were collected from Dera Ghazi Khan district during August till November 2022. The polymerase chain reaction (PCR) analysis amplified a 785 base pair amplicon specific for the Merozoite surface antigen (ms 1-2) gene of T. lestoquardi in 2 out of the 168 (3.3%) sheep blood samples, while no goat blood sample out of 165 was found to be infected with T. lestoquardi. DNA sequencing confirmed the presence of Theileria lestoquardi in both samples and phylogenetic analysis revealed that these amplicon resembled the partial ms 1-2 gene sequences detected in small ruminants from Pakistan, India Iran and Egypt. All the studied epidemiological factors (age, sex, breed, size of herd, dogs with herd, composition of herd, size of herd and Tick burden on sheep) were not found associated with the prevalence of T. lestoquardi. In conclusion, this study reports a low prevalence of T. lestoquardi infection in the Dera Ghazi Khan District of Punjab, Pakistan. The data generated from this work will help pave the way for the prophylactic detection and control of ovine and caprine theileriosis in the region.

Identification and prevalence of ticks infesting small ruminants of Fort Munro region in South Punjab, Pakistan.

Fort Munro is a hill station in Southern Punjab the residents of whom are heavily dependent upon livestock for their living but ticks are a big treat for the livestock. The purpose of this study was to ascertain the tick infestation among the small ruminants of Fort Munro. Ticks (N = 273) were collected from 333 animals (165 goats and 168 sheep) during August and September 2022. Two tick genera Hyalomma and Rhipicephalus were identified. Rhipicephalus was most abundant tick genera (78.02%) followed by Hyalomma (21.98%).Tick distribution significantly varied with the host: Rhipicephalus ticks were more frequently infesting goats while Hyalomma were more common on sheep. For both hosts, tick infestation varied with the sampling sites and male animals were significantly more infested than females. Five tick species were infesting goats with the following relative abundance: Rhipicephalus (R.) senegalensis (39.20%) > R. sanguineus (36.8%) > R. appendiculatus (10.40%) > R. turanicus (8%) > R. guilhoni (5.6%). Rhipicephalus senegalensis (41.22%) was also the most common tick species infesting sheep followed by Hyalomma (H.) marginatum (29.73%), R. guilhoni (18.24%), H. dromedarii (5.41%) and H. impeltatum (5.41%). Male ticks very more abundantly on both goats and sheep than female. Distribution of Hyalomma species also varied between the sampling sites. In conclusion, we are reporting the infestation of 3 Hyalomma and 5 Rhipicephalus in small ruminants of Fort Munro. The data generated through this study will help in developing appropriate tick control in the study area and will add to the existing knowledge regarding tick species that are infesting the small ruminants of Pakistan.

Ovine dicrocoeliosis in Castellón province, eastern Spain: Prevalence, dynamics, and comparative study of different diagnostic techniques.

Dicrocoeliosis is a parasitic disease that mainly affects ruminants during grazing, caused by trematodes of the genus Dicrocoelium, with D. dendriticum being the most common species worldwide. This parasitosis is a chronic and generally subclinical process, with nonspecific signs, which makes its diagnosis challenging. This study aimed to determine the prevalence and seasonal dynamics of D. dendriticum infection in adult sheep from the Valencian Community, eastern Spain, as well as to evaluate the efficacy of flotation and sedimentation techniques when compared with the macroscopic exam of the liver. From February to May 2018, 2019, 2020, 2021, and 2022, a total of 290 adult sheep were examined. The animals were sourced from Castellón province, where a semi-intensive production system predominates. Each animal was euthanized and underwent a macroscopic examination of the liver, as well as a fecal analysis. Among the sampled animals, 117 (40.6%) tested positive for adult trematodes in their liver, while 87 (30%) showed evidence of trematode eggs in the coprological exam, reaching a total of 126 sheep parasitized, with a prevalence of 43.4%. The parasitic burden was established in 90.3 adults per animal when the liver was examined and in 54.5 eggs per gram when the sedimentation coprological exam was performed. No eggs were observed when the flotation technique was employed. A positive correlation was found between the number of adults in the liver and the fecal egg count. No significant differences were detected in the prevalence or parasitic burden throughout the study. Considering the difficulty in controlling the intermediate hosts and the complexity of the life cycle, effective diagnostic methods, combined with the adoption of other preventive measures, is crucial to achieving proper management of this parasitic disease.

Frequency of Besnoitia besnoiti and Neospora caninum antibodies in cattle and small ruminants from greater Cairo and Beni Suef governorates, Egypt.

Livestock industry is a pivotal sector for providing food, work places and monetary support for Egyptian people. Besnoitia besnoiti and Neospora caninum are protozoan parasites that are responsible for significant economic losses in ruminants, particularly in cattle. Besnoitia besnoiti can cause fertility problems and a general loss in productivity, while N. caninum is a major cause of abortion and neonatal abnormalities in infected animals. There is little information on the existence of these protozoa in Egypt, thus we conducted this study to reveal the current situation in cattle (n = 264), sheep (n = 151), and goats (n = 25). Serum samples were collected from governorates of Cairo, Giza, and Beni Suef, representing the most densely populated regions in Egypt. Using commercial ELISAs, an overall estimation among all tested animals (n = 440) revealed 7.7%, 13.2%, and 0.9% as seropositive rates for B. besnoiti, N. caninum, and mixed infection, respectively. Animal species (cattle vs sheep vs goat) and age of cattle (less than vs >1 year old) were analyzed as risk factors for infection. Regarding B. besnoiti, the seroprevalence was significantly higher in cattle than in sheep and goats and in adult cattle than calves. For N. caninum infection, no significant differences were recorded, although the seropositive rates were higher in cattle, and in adult cattle. This study provides the first seroprevalence data for B. besnoiti in all surveyed animals in the regions included, and in sheep and goats from Egypt, and supports the current knowledge for the occurrence of N. caninum in Egypt.

Prevalence and risk factors for gastrointestinal parasitosis in small ruminant farms in the Sahelian zone of Niger.

The general aim of this study is to analyse the risk factors for gastrointestinal parasitosis in small ruminants in order to contribute to the emergence of targeted treatment methods, at herd and agro-climatic zone levels, for the integrated and sustainable management of parasitic diseases in Sahelian livestock systems. The methodology was based on a questionnaire survey conducted in 37 villages and coprological analysis using the McMaster method on faecal samples from 968 small ruminants, including 555 goats and 413 sheep. Multiple logistic regression was used to highlight the risk factors associated with each type of parasitosis encountered. The results showed that the most widespread farming system remained 100% traditional, with feeding based essentially on natural grazing. Coprological results showed the prevalence of nematodosis (70.2%), Cestodosis (4.1%) and Coccidiosis (79.9%), with an average prevalence of coinfection of 56.9%. These parasite loads were significantly higher during the rainy season and in the more arid northern Sahelian zone, with a marked reduction at the end of the season. Average parasitic egg excretions were 1089 EPG of nematodes and 6864 EPG of coccidia. Parasite loads were higher in the wetter southern strip and varied significantly by breed. Of the five breeds of small ruminants studied, the ara-ara sheep had the highest parasitic loads and prevalences for nematodosis (78.6%), coccidiosis (89,3%) and coinfection (70.9%), appears to be the most susceptible to parasitosis. As for risk factors for severe parasite pressure, animals at the end of the rainy season, older animals and those with poor body condition were at risk of nematodiasis or coinfection. On the other hand, animals at the beginning of the rainy season, farms located in less arid southern Sahelian zones and male subjects were the groups at significant risk of coccidiosis. In these extensive Sahelian farming conditions, the control of these parasitoses by selective treatment of animals could be developed, targeting in particular the risk groups highlighted in this study.

Unveiling the presence and genotypic diversity of Giardia duodenalis on large-scale sheep farms: insights from the Henan and Ningxia Regions, China.

BACKGROUND: The parasitic protozoan Giardia duodenalis is an important cause of diarrheal disease in humans and animals that can be spread by fecal-oral transmission through water and the environment, posing a challenge to public health and animal husbandry. Little is known about its impact on large-scale sheep farms in China. In this study we investigated G. duodenalis infection of sheep and contamination of the environment in large-scale sheep farms in two regions of China, Henan and Ningxia. METHODS: A total of 528 fecal samples, 402 environmental samples and 30 water samples were collected from seven large-scale sheep farms, and 88 fecal samples and 13 environmental samples were collected from 12 backyard farms. The presence of G. duodenalis was detected by targeting the ß-giardin (bg) gene, and the assemblage and multilocus genotype of G. duodenalis were investigated by analyzing three genes: bg, glutamate dehydrogenase (gdh) and triphosphate isomerase (tpi). RESULTS: The overall G. duodenalis detection rate was 7.8%, 1.4% and 23.3% in fecal, environmental and water samples, respectively. On the large-scale sheep farms tested, the infection rate of sheep in Henan (13.8%) was found to be significantly higher than that of sheep in Ningxia (4.2%) (P < 0.05). However, the difference between the rates of environmental pollution in Henan (1.9%) and Ningxia (1.0%) was not significant (P > 0.05). Investigations of sheep at different physiological stages revealed that late pregnancy ewes showed the lowest infection rate (1.7%) and that young lambs exhibited the highest (18.8%). Genetic analysis identified G. duodenalis belonging to two assemblages, A and E, with assemblage E being dominant. A total of 27 multilocus genotypes were identified for members of assemblage E. CONCLUSIONS: The results suggest that G. duodenalis is prevalent on large-scale sheep farms in Henan and Ningxia, China, and that there is a risk of environmental contamination. This study is the first comprehensive examination of the presence of G. duodenalis on large-scale sheep farms in China. Challenges posed by G. duodenalis to sheep farms need to be addressed proactively to ensure public health safety.

Moniezia benedeni drives the SNAP-25 expression of the enteric nerves in sheep's small intestine.

BACKGROUND: The neuroimmune network plays a crucial role in regulating mucosal immune homeostasis within the digestive tract. Synaptosome-associated protein 25 (SNAP-25) is a presynaptic membrane-binding protein that activates ILC2s, initiating the host's anti-parasitic immune response. METHODS: To investigate the effect of Moniezia benedeni (M. benedeni) infection on the distribution of SNAP-25 in the sheep's small intestine, the recombinant plasmid pET-28a-SNAP-25 was constructed and expressed in BL21, yielding the recombinant protein. Then, the rabbit anti-sheep SNAP-25 polyclonal antibody was prepared and immunofluorescence staining was performed with it. The expression levels of SNAP-25 in the intestines of normal and M. benedeni-infected sheep were detected by ELISA. RESULTS: The results showed that the SNAP-25 recombinant protein was 29.3 KDa, the titer of the prepared immune serum reached 1:128,000. It was demonstrated that the rabbit anti-sheep SNAP-25 polyclonal antibody could bind to the natural protein of sheep SNAP-25 specifically. The expression levels of SNAP-25 in the sheep's small intestine revealed its primary presence in the muscular layer and lamina propria, particularly around nerve fibers surrounding the intestinal glands. Average expression levels in the duodenum, jejunum, and ileum were 130.32 pg/mg, 185.71 pg/mg, and 172.68 pg/mg, respectively. Under conditions of M. benedeni infection, the spatial distribution of SNAP-25-expressing nerve fibers remained consistent, but its expression level in each intestine segment was increased significantly (P < 0.05), up to 262.02 pg/mg, 276.84 pg/mg, and 326.65 pg/mg in the duodenum, jejunum, and ileum, and it was increased by 101.06%, 49.07%, and 89.16% respectively. CONCLUSIONS: These findings suggest that M. benedeni could induce the SNAP-25 expression levels in sheep's intestinal nerves significantly. The results lay a foundation for further exploration of the molecular mechanism by which the gastrointestinal nerve-mucosal immune network perceives parasites in sheep.