Characterization of intestinal mononuclear phagocyte subsets in young ruminants at homeostasis and during Cryptosporidium parvum infection.

Introduction: Cryptosporidiosis is a poorly controlled zoonosis caused by an intestinal parasite, Cryptosporidium parvum, with a high prevalence in livestock (cattle, sheep, and goats). Young animals are particularly susceptible to this infection due to the immaturity of their intestinal immune system. In a neonatal mouse model, we previously demonstrated the importance of the innate immunity and particularly of type 1 conventional dendritic cells (cDC1) among mononuclear phagocytes (MPs) in controlling the acute phase of C. parvum infection. These immune populations are well described in mice and humans, but their fine characterization in the intestine of young ruminants remained to be further explored. Methods: Immune cells of the small intestinal Peyer's patches and of the distal jejunum were isolated from naive lambs and calves at different ages. This was followed by their fine characterization by flow cytometry and transcriptomic analyses (q-RT-PCR and single cell RNAseq (lamb cells)). Newborn animals were infected with C. parvum, clinical signs and parasite burden were quantified, and isolated MP cells were characterized by flow cytometry in comparison with age matched control animals. Results: Here, we identified one population of macrophages and three subsets of cDC (cDC1, cDC2, and a minor cDC subset with migratory properties) in the intestine of lamb and calf by phenotypic and targeted gene expression analyses. Unsupervised single-cell transcriptomic analysis confirmed the identification of these four intestinal MP subpopulations in lamb, while highlighting a deeper diversity of cell subsets among monocytic and dendritic cells. We demonstrated a weak proportion of cDC1 in the intestine of highly susceptible newborn lambs together with an increase of these cells within the first days of life and in response to the infection. Discussion: Considering cDC1 importance for efficient parasite control in the mouse model, one may speculate that the cDC1/cDC2 ratio plays also a key role for the efficient control of C. parvum in young ruminants. In this study, we established the first fine characterization of intestinal MP subsets in young lambs and calves providing new insights for comparative immunology of the intestinal MP system across species and for future investigations on host-Cryptosporidium interactions in target species.

The identification of small molecule inhibitors with anthelmintic activities that target conserved proteins among ruminant gastrointestinal nematodes.

Gastrointestinal nematode (GIN) infections are a major concern for the ruminant industry worldwide and result in significant production losses. Naturally occurring polyparasitism and increasing drug resistance that potentiate disease outcomes are observed among the most prevalent GINs of veterinary importance. Within the five major taxonomic clades, clade Va represents a group of GINs that predominantly affect the abomasum or small intestine of ruminants. However, the development of effective broad-spectrum anthelmintics against ruminant clade Va GINs has been challenged by a lack of comprehensive druggable genome resources. Here, we first assembled draft genomes for three clade Va species (Cooperia oncophora, Trichostrongylus colubriformis, and Ostertagia ostertagi) and compared them with closely related ruminant GINs. Genome-wide phylogenetic reconstruction showed a relationship among ruminant GINs structured by taxonomic classification. Orthogroup (OG) inference and functional enrichment analyses identified 220 clade Va-specific and Va-conserved OGs, enriched for functions related to cell cycle and cellular senescence. Further transcriptomic analysis identified 61 taxonomically and functionally conserved clade Va OGs that may function as drug targets for new broad-spectrum anthelmintics. Chemogenomic screening identified 11 compounds targeting homologs of these OGs, thus having potential anthelmintic activity. In in vitro phenotypic assays, three kinase inhibitors (digitoxigenin, K-252a, and staurosporine) exhibited broad-spectrum anthelmintic activities against clade Va GINs by obstructing the motility of exsheathed L3 (xL3) or molting of xL3 to L4. These results demonstrate valuable applications of the new ruminant GIN genomes in gaining better insights into their life cycles and offer a contemporary approach to discovering the next generation of anthelmintics.IMPORTANCEGastrointestinal nematode (GIN) infections in ruminants are caused by parasites that inhibit normal function in the digestive tract of cattle, sheep, and goats, thereby causing morbidity and mortality. Coinfection and increasing drug resistance to current therapeutic agents will continue to worsen disease outcomes and impose significant production losses on domestic livestock producers worldwide. In combination with ongoing therapeutic efforts, advancing the discovery of new drugs with novel modes of action is critical for better controlling GIN infections. The significance of this study is in assembling and characterizing new GIN genomes of Cooperia oncophora, Ostertagia ostertagi, and Trichostrongylus colubriformis for facilitating a multi-omics approach to identify novel, biologically conserved drug targets for five major GINs of veterinary importance. With this information, we were then able to demonstrate the potential of commercially available compounds as new anthelmintics.

Two in vitro anthelmintic assays of four Paraguayan medicinal plants for proof of concept of the role of polyphenols in their biological activities and LC-HRMS analysis.

ETHNOPHARMACOLOGICAL RELEVANCE: Gastrointestinal nematodes (GIN) control in small ruminants has relied on the systematic use of synthetic anthelmintics (AH), their effectiveness has been progressively decreasing due to the rise and diffusion of anthelmintic resistances. The most prevalent genera affecting small ruminants were Haemonchus spp., and Trichostrongylus spp. The investigation of new anthelmintics in plants is a highly studied option, especially when it is linked to ethnobotanical knowledge and phenolic compounds. THE AIMS OF OUR STUDY: Four medicinal plants mentioned in traditional medicine were selected to evaluate their anthelmintic proprieties at different stages of the life cycle of GIN, namely: Kyllinga odorata Valh., Cassia occidentalis L., Artemisia absinthium L, and Verbena litoralis Kunth and to explore the role of polyphenols in the AH activity. MATERIALS AND METHODS: To evaluate the anthelmintic activity in this study, two models of GIN species, namely Haemonchus contortus (Hc) and Trichostrongylus colubriformis (Tc) were selected and tested on two in vitro assays: 1) Larval Exsheathment Inhibition Assay (LEIA) and, 2) Egg Hatch Assay (EHA). To explore the role of tannins and polyphenols in AH activity by comparing the effects of LEIA and EHA with or without polyvinylpolypyrrolidone (PVPP) and to characterize the phytochemical composition of the most active plants using ultra-high performance liquid chromatography (UHPLC) coupled with high-resolution mass spectrometry (HRMS). RESULTS: C. occidentalis exhibited the highest activity on LEIA (EC50 = 250.42-41.80 µg/mL) and A. absinthium on egg hatching processes (EC50 = 121.70-137.34 µg/mL) in both species of GIN. The inhibition in the development of eggs was from 67.70% to 96.36% on H. contortus, and from 78.87% to 99.65% on T. colubriformis. At the maximal dose, Additionally, it was observed that the AH on eggs varies according to the GIN species: on H. contortus the extracts tested blocked the formation of larvae Ovicidal Effect (% higher OE) and on T. colubriformis they blocked the appearance of L1 larvae, Larvae Failing Eclosion (% higher LFE). After PVPP, a reduction in AH activity on LEIA and EHA was noted, especially with C. occidentalis (87.20-67.00% of larvae exsheathment, (p < 0.05) and 40.51-24.96% of egg hatching, (p > 0.05) of both parasite species. Nine putative features were identified using HRMS and MS/MS after addition of PVPP. CONCLUSIONS: The present study demonstrated that C. occidentalis, A. absinthium, and K. odorata, which parts have been traditionally used as medicinal plants are a valuable source of active compounds with anthelmintic activity. The medicinal use of these plants against GIN parasites was proven by in vitro analysis. Therefore exploration of the secondary metabolites of these plant extracts and testing of isolated fractions of active compounds under in vivo experiments are planned and represent a specific challenge for alternative drug research. Regarding the PVPP, in this study we hypotheses about the standard doses it was not able to completely absorb the polyphenols of extracts of K. odorata, C. occidentalis, and A. absinthium, which would lead to more studies to evaluate the role of this product in the absorption of phenolic compounds.

Prevalence of gastrointestinal parasite in small ruminants of District Dir Upper Khyber Pakhtunkhwa Province of Pakistan / Prevalência de parasita gastrointestinal em pequenos ruminantes do distrito Upper Dir, da província de Khyber Pakhtunkhwa, no Paquistão

The present research was planned to assess the occurrence of intestinal parasites in small ruminants of Upper Dir of Khyber Pakhtunkhwa Province of Pakistan. For this purpose, the faecal material was collected randomly with gloved fingers directly from the rectum region of sheep and goats and the faecal materials were then put in hygienic plastic bottles with 10% formalin. The overall 315 (n=184 sheep and n= 131 goats) faecal samples were collected out of 315 samples, 281 were found positive for different parasites. Patterns-wise prevalence of GI parasites of the study area was found. Overall Single parasitism 89.20% (281/315) with 94.0% (173/184) in sheep and 82.43% (108/131) in goats. Double parasitic infection in small ruminant recorded in which Fasciola+ Haemonchus. contortus in sheep were found their prevalence was 25.54% (47/184). While in goats, the double parasitic infection in which Haemonchus contortus+Trichuris spp were found and their prevalence were 23.43% (30/131). The species found in the sample of sheep were includes, i.e., Strongyloides papillosus (41.30%), Heamonchus controtus (21.73%), Trichuris ovis (17.39%), and Fasciola hepatica (13.58%), the corresponding value for goat were Strongyloides spp 33.33% (36/108), Haemonchus spp 28.70%, (27/108), Trichuris spp 25.20% (27/184) and Fasciola spp 10.68% (14/184). The sheep of the study area are more infected as compared to goats. This study suggested that gastrointestinal parasites are major health problems of small ruminants in the study area. Therefore, a comprehensive study on species of gastrointestinal parasites circulating in the area, control options, cost-effective strategies and awareness about gastrointestinal parasites among the farmers in the study area should be instituted.

A presente pesquisa foi planejada para avaliar a ocorrência de parasitas intestinais em pequenos ruminantes do distrito Upper Dir, da província de Khyber Pakhtunkhwa, no Paquistão. Para tanto, o material fecal foi coletado aleatoriamente com dedos enluvados diretamente da região do reto de ovelhas e cabras, e os materiais fecais foram colocados em garrafas plásticas higiênicas com formol a 10%. No total, 315 (n = 184 ovelhas e n = 131 cabras) amostras fecais foram coletadas; destas, 281 foram consideradas positivas para diferentes parasitas. A prevalência de padrões de parasitas GI da área de estudo foi encontrada. Parasitismo global único 89,20% (281/315) com 94,0% (173/184) em ovinos e 82,43% (108/131) em cabras. Infecção parasitária dupla em pequenos ruminantes registrada em Fasciola + Haemonchus contortus em ovinos, sua prevalência foi de 25,54% (47/184). Já em caprinos, a dupla infecção parasitária em que Haemonchus contortus + Trichuris spp foram encontrados e sua prevalência foi de 23,43% (30/131). As espécies encontradas na amostra de ovinos foram: Strongyloides papillosus (41,30%), Heamonchus controtus (21,73%), Trichuris ovis (17,39%) e Fasciola hepatica (13,58%), o valor correspondente para cabra foi Strongyloides spp. 33,33% (36/108), Haemonchus spp. 28,70%, (27/108), Trichuris sp 25,20% (27/184) e Fasciola spp. 10,68% (14/184). As ovelhas da área de estudo estão mais infectadas do que as cabras. Este estudo sugeriu que os parasitas gastrointestinais são os principais problemas de saúde dos pequenos ruminantes na área de estudo. Portanto, um estudo abrangente sobre as espécies de parasitas gastrointestinais que circulam na área, opções de controle, estratégias de baixo custo e conscientização sobre parasitas gastrointestinais entre os agricultores na área de estudo deve ser instituído.

Immune reactivity and host modulatory roles of two novel Haemonchus contortus cathepsin B-like proteases.

BACKGROUND: Haemonchus contortus is a blood-feeding, gastrointestinal nematode (GIN) that causes significant economic losses to the small ruminant industry worldwide. Despite extensive efforts, our understanding of the molecular mechanisms used by GIN to evade host immune responses is limited. Cathepsin B-like proteins (CBPs) are members of the cysteine protease family and are involved in parasite invasion and thus provide viable vaccine candidates. METHODS: In silico comparative analysis was used to identify conserved proteins among a subset of clade V parasitic nematodes with emphasis on blood-feeding worms, among which CBPs appeared prominently. We identified and characterized two novel CBPs designated Hc-CBP-1 and Hc-CBP-2. Rabbit anti-recombinant (r) Hc-CBP-1 and rHc-CBP-2 were used to detect the presence of native proteins in the excretory secretory products (ESP) and in worm tissues of adult H. contortus. Peptide arrays of rHc-CBP-1 and rHc-CBP-2 were screened with the homologous and heterologous anti-sera and with sera from dexamethasone-treated (Dex+) and non-treated (Dex-) H. contortus-infected animals to identify key immunogenic peptides. Gene transcription of Hc-cbp-1 and Hc-cbp-2 was also performed on H. contortus-infected animals treated with Dex+. Finally, the mature recombinant proteins were used to assess their abilities to modulate cell functions. RESULTS: Immunohistochemistry showed that both Hc-CBP-1 and Hc-CBP-2 are present on the brush borders of the intestine; Hc-CBP-2 was also present in the hypodermis of the body wall. Peptide displays screened with rabbit anti-rHc-CBP-1 and anti-rHc-CBP-2 revealed regions within the proteins where dominant and overlapping epitopes prevailed. ELISA results were consistent with only Hc-CBP-1 being present in H. contortus adult ESPs. H. contortus from Dex+ animals exhibited a threefold increase in Hc-cbp-2 transcript while Hc-cbp-1 expression did not change. In contrast, comparisons of immunoreactivities of rHc-CBP-1 and rHc-CBP-2 peptide arrays to sera from Dex+ and Dex- animals primarily showed changes in Hc-CBP-1 binding. Lastly, rHc-CBP-1 suppressed mRNA expression of bovine peripheral blood mononuclear cell cytokines/activation markers, including TNFα, IL-1, IL-6 and CD86. CONCLUSIONS: These results suggest that as secreted and cryptic proteins, respectively, Hc-CBP-1 and Hc-CBP-2 influence cellular and immunological activities that have interesting dynamics during infection and may provide viable immune-related targets for attenuating H. contortus infectivity.

Seroepidemiology of Toxoplasma gondii in wild ruminants in Spain.

Toxoplasmosis is a parasitic zoonosis caused by Toxoplasma gondii which infects warm-blooded species worldwide. Humans can be infected through ingestion of tissue cysts from raw or undercooked meat, including game meat. A nationwide large-scale cross-sectional study was conducted to assess exposure to T. gondii in seven wild ruminant species in Spain. A total of 2,040 serum samples from 77 sampling sites randomly distributed in the five bioregions (BRs) covering mainland Spain were tested for antibodies against T. gondii using the modified agglutination test. The overall seroprevalence was 22.0% (449/2,040). Seroprevalence by species in decreasing order was as follows: 39.6% (141/356) in roe deer (Capreolus capreolus), 37.1% (138/372) in fallow deer (Dama dama), 16.6% (92/553) in red deer (Cervus elaphus), 14.0% (26/186) in Southern chamois (Rupicapra pyrenaica), 11.5% (24/209) in mouflon (Ovis aries musimon), 7.8% (27/346) in Iberian wild goat (Capra pyrenaica) and 5.6% (1/18) in Barbary sheep (Ammotragus lervia). Seropositivity was detected in 74.0% (57/77) of the sampling sites. Results indicate widespread but not homogeneous exposure to T. gondii in wild ruminant populations in Spain during the last two decades and highlight differences related to animal species and spatial distribution of these species in this country; this implies potential consequences of T. gondii for animal health, conservation and public health.

Epidemiological approach to nematode polyparasitism occurring in a sympatric wild ruminant multi-host scenario.

The epidemiology behind multi-host/multi-parasite systems is particularly interesting to investigate for a better understanding of the complex dynamics naturally occurring in wildlife populations. We aimed to approach the naturally occurring polyparasitism of gastrointestinal nematodes in a sympatric wild ruminant scenario present in south-east Spain. To this end, the gastrointestinal tract of 252 wild ruminants of four different species (red deer, Cervus elaphus; mouflon, Ovis aries musimon; Iberian ibex, Capra pyrenaica and fallow deer, Dama dama) were studied in Cazorla, Segura y Las Villas Natural Park (Andalusia, Spain). Of the analysed animals, 81.52% were positive for parasite infection and a total of 29 nematode species were identified. Out of these, 25 species were detected in at least two host species and 11 parasitized all ruminant species surveyed. The multi-host interaction between these nematodes and the four host species is discussed under the perspective of host family-based differences.

Seroprevalence of Toxoplasma gondii and associated hematological alterations in small ruminants of D.G. Khan district of Southern Punjab, Pakistan / [Soroprevalência de Toxoplasma gondii e alterações hematológicas associadas em pequenos ruminantes no distrito de D.G. Khan no sul de Punjab, Paquistão]

The present study was carried out to evaluate the prevalence and hematological effects of Toxoplasma gondii in sheep and goat in district Dera Ghazi Khan. Blood samples (n=204) were collected comprise goats (n=101) and sheep (n=103) alongwith age, gender and breeds of animals. Samples were collected randomly from 25 flocks of 7 different union council Viz. Vehova, Tibbi Qaisrani, Lakhani, Kohar, Tuman Qaisrani, Nutkani and Kot Qaisrani of Tehsil Taunsa Sharif at least 4 animals from each flock. All ruminants divide into three groups based on age, breed and gender. The prevalence was detected through two different kits Viz. LAT and ELISA kit. The overall prevalence suspected in goats through LAT and ELISA kit was (35.64%), (32.67%) and in sheep was (25.24%), (23.30%) respectively. The Toxoplasma gondii had a significant effect on goats in age groups and non-significant all other groups of goats and sheep. Toxoplasma gondii had a significant effect on all hematological parameters like Hemoglobin, total leukocyte cells, granulocytes, lymphocytes, platelets, and red blood cells, except monocytes. In conclusion of the current study, toxoplasmosis is prevalent among ruminants, reveals the possibility of transmission to humans on the use of host animals as protein source.(AU)

O objetivo do presente estudo foi avaliar a prevalência e efeitos hematológicos de Toxoplasma gondii em ovelhas e cabras no distrito Dera Ghazi Khan. Amostras de sangue (n=204) foram coletadas para incluir cabras (n=101) e ovelhas (n=103), além de idade, gênero e raça dos animais. Amostras foram coletadas aleatoriamente de 25 rebanhos de 7 conselhos sindicais: Vehova, Tibbi Qaisrani, Lakhani, Kohar, Tuman Qaisrani, Nutkani e Kot Qaisrani of Tehsil Taunsa Sharif com pelo menos 4 animais em cada rebanho. Todos os ruminantes foram divididos em três grupos baseados em idade, raça e gênero. A prevalência foi detectada usando dois kits, LAT e ELISA. A prevalência total suspeita em cabras através dos kits LAT e ELISA foi (35.64%), (32.67%) e em ovelhas foi (25.24%), (23.30%) respectivamente. O Toxoplasma gondii teve efeito significativo em cabras em grupos de idade e não significativo em todos os outros grupos de cabras e ovelhas. Toxoplasma gondii teve efeito significativo em todos os parâmetros hematológicos como hemoglobina, células totais de leucócitos, granulócitos, linfócitos, plaquetas e glóbulos vermelhos, exceto monócitos. O presente estudo conclui que toxoplasmose é prevalente entre ruminantes, e revela a possibilidade de transmissão para humanos com o uso de animais hospedeiros como fonte de proteína.(AU)

Neosporosis, Toxoplasmosis, and Sarcocystosis in Ruminants: An Update.

Neospora caninum, Toxoplasma gondii, and Sarcocystis spp. are related Apicomplexan parasites that have 2 hosts in their life cycles. The definitive hosts excrete unsporulated (Neospora caninum, T gondii) or sporulated (Sarcocystis spp.) oocysts in their feces after ingesting tissue cysts from the tissues of ruminant intermediate hosts. These coccidians can cause abortion and neonatal mortality in ruminants. T gondii and Sarcocystis hominis (from cattle) are zoonotic. This article reviews information on the etiology, life cycle, diagnosis, control and prevention of these parasites and the diseases they cause in ruminants.

Species-specific differences in Toxoplasma gondii, Neospora caninum and Besnoitia besnoiti seroprevalence in Namibian wildlife.

BACKGROUND: Knowledge about parasitic infections is crucial information for animal health, particularly of free-ranging species that might come into contact with livestock and humans. METHODS: We investigated the seroprevalence of three tissue-cyst-forming apicomplexan parasites (Toxoplasma gondii, Neospora caninum and Besnoitia besnoiti) in 506 individuals of 12 wildlife species in Namibia using in-house enzyme linked immunosorbent assays (indirect ELISAs applying purified antigens) for screening and immunoblots as confirmatory tests. We included six species of the suborder Feliformia, four species of the suborder Caniformia and two species of the suborder Ruminantia. For the two species for which we had most samples and life-history information, i.e. cheetahs (Acinonyx jubatus, n = 250) and leopards (Panthera pardus, n = 58), we investigated T. gondii seroprevalence in relation to age class, sex, sociality (solitary, mother-offspring group, independent sibling group, coalition group) and site (natural habitat vs farmland). RESULTS: All but one carnivore species (bat-eared fox Otocyon megalotis, n = 4) were seropositive to T. gondii, with a seroprevalence ranging from 52.4% (131/250) in cheetahs to 93.2% (55/59) in African lions (Panthera leo). We also detected antibodies to T. gondii in 10.0% (2/20) of blue wildebeest (Connochaetes taurinus). Adult cheetahs and leopards were more likely to be seropositive to T. gondii than subadult conspecifics, whereas seroprevalence did not vary with sex, sociality and site. Furthermore, we measured antibodies to N. caninum in 15.4% (2/13) of brown hyenas (Hyaena brunnea) and 2.6% (1/39) of black-backed jackals (Canis mesomelas). Antibodies to B. besnoiti were detected in 3.4% (2/59) of African lions and 20.0% (4/20) of blue wildebeest. CONCLUSIONS: Our results demonstrate that Namibian wildlife species were exposed to apicomplexan parasites at different prevalences, depending on parasite and host species. In addition to serological work, molecular work is also needed to better understand the sylvatic cycle and the clear role of wildlife in the epidemiology of these parasites in southern Africa.

In vitro anthelmintic activity of Lippia gracilis Schauer essential oil against egg-hatching of goat gastrointestinal nematodes / Atividade anti-helmíntica in vitro do óleo essencial de Lippia gracilis Schauer contra a eclosão de nematódeos gastrintestinais de caprinos

Since drug-resistant nematodes became a common problem in sheep and goat industries, alternative methods using natural products have emerged as a viable and sustainable anthelmintic treatment option. Here, the in vitro effect of essential oil extracted from Lippia gracilis Schauer was assessed on the hatching process of nematodes recovered from naturally infected goats. Essential oil at concentrations of 0.08% (0.008 µL/mL), 0.12% (0.012 µL/mL), and 0.16% (0.016 µL/mL) was able to induce an average inhibition of 74.7, 84 and 93%, respectively. The effective concentration required to inhibit egg hatching in 50% of eggs (EC50) was 0.03452%. Therefore, essential oil of L. gracilis showed promisor in vitro anthelmintic results against egg-hatching of goat gastrointestinal nematodes.(AU)

Como os nematoides resistentes a drogas se tornaram um problema comum nas indústrias de ovinos e caprinos, métodos alternativos que utilizam produtos naturais surgiram como uma opção de tratamento anti-helmíntico viável e sustentável. Aqui, o efeito in vitro do óleo essencial extraído de Lippia gracilis Schauer foi avaliado no processo de eclosão de nematoides recuperados de caprinos naturalmente infectadas. O óleo essencial nas concentrações de 0,08% (0,008 µL/mL), 0,12% (0,012 µL/mL), e 0,16% (0,016 µL/mL)foi capaz de induzir uma inibição média de 74,7, 84 e 93%, respectivamente. A concentração efetiva necessária para inibir a eclosão de ovos em 50% dos ovos (CE50) foi de 0,03452%. Portanto, o óleo essencial de L. gracilis apresentou resultados anti-helmínticos in vitro promissores contra a eclosão de nematódeos gastrintestinais de caprinos.(AU)

Potential of botanical driven essential oils against Haemonchus contortus in small ruminants / Potencial de aceites esenciales botánicos contra Haemonchus contortus en pequeños rumiantes

The livestock sector is continuously facing problems in controlling parasitic diseases especially Haemonchosis due to emergence of anthelminthic resistance and failure in vaccination control programmes. Therefore, to increase milk and meat production and emerging demand of meat free from drug residues development of new alternative approaches are appealing for prevention and control of Haemonchosis in small ruminants. Among alternatives, plants driven essentials oils have shown promising results in control of Haemonchus contortus infection at various concentrations by different assays including egg hatch assay, larval development assay, larval exsheathment assay and adult motility assay. Essential oils are complex mixtures of various impulsive or volatile compounds which have potential to control Haemonchosis. The current study reviews the therapeutic effects of essential oils of plants against Haemonchus contortus and to be used them against Haemonchus contortus for future perspectives.

El sector ganadero enfrenta continuamente problemas para controlar las enfermedades parasitarias, especialmente la hemoncosis, debido a la aparición de resistencia antihelmíntica y al fracaso en los programas de control de vacunación. Por lo tanto, para aumentar la producción de leche y carne, y la demanda emergente de carne libre de residuos de medicamentos, el desarrollo de nuevos enfoques alternativos es atractivo para la prevención y el control de la hemoncosis en pequeños rumiantes. Entre las alternativas, los aceites esenciales producidos por las plantas han mostrado resultados prometedores en el control de la infección por Haemonchus contortus a diversas concentraciones mediante diferentes ensayos, incluido el análisis de eclosión de huevos, el desarrollo de larvas, el análisis de vaciado de larvas y el ensayo de motilidad en adultos. Los aceites esenciales son mezclas complejas de varios compuestos impulsivos o volátiles que tienen potencial para controlar la hemonchosis. Este estudio revisa los efectos terapéuticos de los aceites esenciales de las plantas contra Haemonchus contortus y evalúa sus perspectivas futuras como agentes para combatir las enfermedades causadas por este parásito.

Parasitos gastrintestinais em caprinos no município de Quixadá, Ceará / Gastrointestinal parasites in goats from Quixadá, Ceará

O objetivo desse trabalho foi avaliar a ocorrência dos principais parasitos gastrintestinais que acometem caprinos adultos e jovens do município de Quixadá-Ceará durante a época seca e chuvosa, além de identificar e quantificar as espécies de Eimeria presentes no rebanho. Foram utilizados 654 animais, sendo 334 animais no período seco, dos quais eram 149 matrizes e 185 crias e 320 animais no período chuvoso, sendo 106 matrizes e 214 crias, independente de padrão racial e sistema de criação. As coletas foram realizadas nos meses de novembro e dezembro de 2012 e maio de 2013 no município de Quixadá-Ceará e encaminhadas ao Laboratório de Parasitologia da Embrapa Caprinos e Ovinos na cidade de Sobral-Ceará. Foram coletadas fezes para contagem do número de ovos por gramas de fezes (OPG), contagem de Oocistos por gramas de fezes (OOPG), coprocultura e identificação das espécies de Eimeria com base na morfometria. As matrizes apresentaram maiores médias de ovos do tipo Strongyloidea (372,03) e Strongyloides (8,45) no período seco e no período chuvoso apresentaram maiores médias apenas para Strongyloidea (502,85). O inverso aconteceu com o OOPG, onde as crias apresentaram maiores médias no período seco (9387,41) e no chuvoso (9630,37). O gênero de nematódeo mais frequente na coprocultura foi o Trichostrongylus sp. (63,67%) no período seco e o Haemonchus sp. no chuvoso (66,67%). Em 100% dos rebanhos, foram encontrados oocistos de Eimeria, com 81% dos animais infectados. A ocorrência da eimeriose nas categorias jovens e adultas foi de 64,3% e 83,2% no período seco e de 87,4% e 92,4% no período chuvoso. As espécies identificadas foram: Eimeria alijevi, E. apsheronica, E. arloingi, E. caprina, E. caprovina, E. christenseni, E. hirci, E. jolchijevi e E. ninakohlyakimovae. Nas crias a E. alijevi foi a mais freqüente no período seco e a E. ninakohlyakimovae no chuvoso. Já nas matrizes a espécie E. alijevi apresentou a maior frequência em ambos os períodos. Nos rebanhos estudados foi evidenciado a presença de ovos de nematódeos gastrointestinais e oocistos de Eimeria spp. independente da categoria animal.(AU)

The objective of this study was to evaluate the occurrence of main gastrointestinal parasites and to quantify and identify the species of Eimeria present in young goats and adults in the city of Quixadá, Ceará, during the dry and rainy season. Six hundred fifty-four animals were used, with 334 animals during the dry season, which were 149 mothers and 185 offspring and 320 animals during the rainy season, with 106 mothers and 214 offspring, independent of breed standard and creation system. Samples were collected in the months of November and December 2012 and May 2013 in the city of Quixadá, Ceará and sent to the Parasitology Laboratory of Embrapa Goats and Sheep at Sobral, Ceará, feces were collected for counting the number of eggs per gram of feces (EPG), counting oocysts per gram of feces (OOPG), coproculture and identification of Eimeria species based on the morphometry. Matrices showed higher means egg type Strongyloidea (372.03) and Strongyloides (8.45) in the dry season and the rainy season had higher mean only to Strongyloidea (502.85). The inverse happened to OOPG, where the offspring showed higher means in the dry season (9387.41) and rainy (9630.37). The genus most frequent of nematode in coprocultures was Trichostrongylus. (63.67%) in the dry season and Haemonchus sp. the rainy (66.67%). One hundred percent of the herds, oocysts of Eimeria were found, with 81% of infected animals. The occurrence of eimeriosis in young and adult categories was 64.3% and 83.2% in the dry season and 87.4% and 92.4% during the rainy season. The species identified were: Eimeria alijevi, E. apsheronica, E. arloingi, E. caprina, E. caprovina, E. christenseni, E. hirci, E.and E. jolchijevi ninakohlyakimovae. In the offspring E. alijevi was the most frequent in the dry season and the rainy E. ninakohlyakimovae. Already in matrices the species E. Alijevi showed the highest frequency in both periods. In herds studied it evidenced the presence of gastrointestinal nematodes eggs and oocysts of Eimeria spp. independent of animal category.(AU)

Prevalence and staining characteristics of Blastocystis isolates from food animals in Tamil Nadu.

The present study was undertaken to investigate the prevalence and staining characteristics of Blastocystis isolated from food animals. Smears of the duodenal and caecal mucosal scrapings, collected from food animals, were stained with Giemsa, Gram's, modified acid-fast and acridine orange. Blastocystis was identified in 295 samples, including faeces and intestinal contents of animals like small ruminants (95), poultry (170) and pigs (30). The prevalence in pigs was found to be high (94.4%) followed by poultry (29.4%) and small ruminants (14%). Various forms of Blastocystis such as vacuolar, granular and amoeboid forms were identified by using different stains. The parasites stained with Giemsa were identified by the presence of eosinophilic nucleus and basophilic cytoplasm. In organisms stained with Gram's stain, the cytoplasm of the vacuolar forms took up the counter stain safranine. Blastocystis appeared as a pink colored cyst against bluish green background with modified acid-fast staining. The study shows that there is a very high prevalence of Blastocystis among the food animals investigated. Simple parasitological procedures, including direct microscopical examination and staining with agents like Giemsa, Gram's and acridine orange can assist identification of the parasites from intestinal contents and faecal material.

Innate immune responses play a key role in controlling infection of the intestinal epithelium by Cryptosporidium.

Cryptosporidium infection leads to acute diarrhea worldwide. The development of cryptosporidiosis is closely related to the immune status of its host, affecting primarily young ruminants, infants, and immunocompromised individuals. In recent years, several studies have improved our knowledge on the immune mechanisms responsible for the control of the acute phase of the infection and have highlighted the importance of innate immunity. The parasite develops in the apical side of intestinal epithelial cells, giving these cells a central role, as they are both the exclusive host cell for replication of the parasite and participate in the protective immune response. Epithelial cells signal the infection by producing chemokines, attracting immune cells to the infected area. They also actively participate in host defense by inducing apoptosis and releasing antimicrobial peptides, free or incorporated into luminal exosomes, with parasiticidal activity. The parasite has developed several escape mechanisms to slow down these protective mechanisms. Recent development of several three-dimensional culture models and the ability to genetically manipulate Cryptosporidium will greatly help to further investigate host-pathogen interactions and identify virulence factors. Intestinal epithelial cells require the help of immune cells to clear the infection. Intestinal dendritic cells, well known for their ability to induce and orchestrate adaptive immunity, play a key role in controlling the very early steps of Cryptosporidium parvum infection by acting as immunological sentinels and active effectors. However, inflammatory monocytes, which are quickly and massively recruited to the infected mucosa, seem to participate in the loss of epithelial integrity. In addition to new promising chemotherapies, we must consider stimulating the innate immunity of neonates to strengthen their ability to control Cryptosporidium development. The microbiota plays a fundamental role in the development of intestinal immunity and may be considered to be a third actor in host-pathogen interactions. There is an urgent need to reduce the incidence of this yet poorly controlled disease in the populations of developing countries, and decrease economic losses due to infected livestock.

Paleoparasitological analysis of the extinct Myotragus balearicus Bate 1909 (Artiodactyla, Caprinae) from Mallorca (Balearic Islands, Western Mediterranean).

Myotragus balearicus (Artiodactyla, Caprinae) is an extinct caprine endemic of the Eastern Balearic Islands or Gymnesics (i.e., Mallorca, Menorca and surrounding islets, Western Mediterranean Sea). In spite of its small size, c. 50cm height at the shoulder, it was the largest mammal inhabiting these islands until the human arrival, and it had peculiar short legs and frontal vision. It disappeared between 2830 and 2210calBCE. The coprolites here studied were recovered from Cova Estreta, in Pollença, Mallorca. The samples were subjected to microscopic examination and enzyme-linked immunosorbent assays (ELISA) for E. histolytica/E. dispar, Giardia intestinalis and Cryptosporidium parvum. This study provides new paleoparasitological data from an extinct animal species of the Holocene period. The microscopy revealed one sample containing uninucleated-cyst of Entamoeba sp., whereas ELISA detected nine positive samples for Cryptosporidium sp. The finding of these protozoans can help in the discussion of its extinction cause and demonstrates the antiquity and the evolutionary history of host-parasite relationships between protozoa and caprines since the Messinian.

Quitosana fúngica sobre larvas de nematoides gastrintestinais de caprinos / Fungal chitosan on gastrointestinal nematodes larvae of goats

A caprinocultura é representada por um efetivo bastante considerável no Nordeste brasileiro, porém, infecções causadas por nematoides e o sério problema da resistência parasitária se tornaram barreiras para a criação desses animais. Como alternativa, o controle com bioprodutos entra como uma solução sustentável e viável para auxiliar na criação da região. Nesse contexto, o presente trabalho avaliou a atuação da quitosana fúngica sobre o desenvolvimento larval de nematoides gastrintestinais em amostras de caprinos naturalmente infectados. Para tanto, foi realizada a seleção de 5 propriedades e confirmada a positividade do rebanho, além de coproculturas com solução de quitosana a 0,5; 1,0 e 1,5%, com cada tratamento realizado em 5 repetições. As larvas de terceiro estágio (L3) foram recuperadas e cem larvas por tratamento foram contabilizadas e identificadas. Os gêneros identificados foram Haemonchus, Strongyloides, Oesophagostomum e Trichostrongylus. Na análise da inibição do desenvolvimento larval, a concentração de 1,0% impediu o desenvolvimento larval do Haemonchus em 35%, porém, os resultados não tiveram diferença estatística significante. Assim, sugere-se buscar novas concentrações de quitosana fúngica como anti-helmíntico, visto que se apresenta como uma alternativa promissora no controle sustentável desses endoparasitos.(AU)

The goat is represented by a very considerable effective in the Northeastern Brazil, but infections caused by nematodes and the serious problem of parasitic resistance have become barriers to breed these animals. Alternatively, the control with bioproducts comes as a sustainable and viable solution to help breeding in this region. In this context, the present study evaluated the performance of fungal chitosan on the larval development of gastrointestinal nematodes in naturally infected goat samples. Therefore, the selection was performed at five properties. The positive herd was confirmed, and coprocultures were performed with chitosan solution 0.5, 1.0 and 1.5%, with each treatment performed in 5 replicates. The third-stage larvae (L3) were recovered and one hundred larvae/treatment were counted and identified. The identified genera were Haemonchus, Strongyloides, Oesophagostomum and Trichostrongylus. In the analysis of inhibition of larval development, the concentration of 1.0% prevented the development of larval Haemonchus by 35%, but the results were not statistically significant. Thus, it is suggested to seek new concentrations of fungal chitosan as anthelmintic, since it appears as a promising alternative to sustainable control of these endoparasites.(AU)

Systematic review and meta-analysis of metacestodes prevalence in small ruminants in Ethiopia.

Metacestodes, the larval stages of canid cestode parasites, are among the causes of morbidity, mortality and financial losses in small ruminants in Ethiopia as a result of organ and carcass condemnation at slaughter. Several studies have been conducted over the years; however, these studies often had limited scope and coverage. This systematic review and meta-analysis was conducted to collate the information so far available in order to provide a pooled prevalence estimate at national level and identify potential predictors. Published and grey literature written in English and Amharic in the period from, 1st of January 1990 to June 25, 2015 were searched from electronic databases and repositories of academic and research institutions. Relevant animal level data on 67,743 small ruminants was extracted from 23 published articles and one master's thesis resulting altogether in 86 animal level reports that conformed to predefined criteria. The dataset was analyzed using a meta-analytical approach. The pooled prevalence estimate computed for metacestodes infection was 11.8% with a 95% confidence interval (CI) of 9.1, 15.4. The highest prevalence estimate 31.2% (95% CI: 23.1, 40.9) was found for Cysticercus tenuicollis (Taenia hydatigena) followed by cystic echinococcosis (Echinococcus granulosus) 8.8% (95% CI: 5.8, 13.1), Cysticercus ovis (Taenia ovis) 4.9% (95% CI: 2.9, 9.4) and Coenurus cerebralis (Taenia multiceps) 4.6% (95% CI: 1.6, 12.2). Among the predictors considered for heterogeneity analysis only sample size and metacestode type fitted the final multivariable meta-regression model and explained 26.3% of the explainable heterogeneity between studies (p<0.05). The prevalence was noted to decrease with increasing sample size. No significant difference in prevalence was observed between sheep and goats (p>0.05). In conclusion, this review showed a widespread occurrence of metacestodes in small ruminants in Ethiopia. Thus, a holistic approach to break the life cycle of these parasitic stages is suggested, including regulatory interventions that encourage dog owners to keep their dogs confined and prevent backyard slaughter and proper management of abattoir waste disposal.

Molecular characterization of veterinary important trematode and cestode species in the mithun Bos frontalis from north-east India.

Helminth infections in the mithun Bos frontalis, including the liver fluke Fasciola gigantica, hepato-gastric amphistomes Explanatum explanatum, Paramphistomum epiclitum and Calicophoron calicophorum, and the cestodes Echinococcus granulosus and E. ortleppi were studied in north-east India over a 2-year period from 2012 to 2014. Cystic echinococcosis caused by E. granulosus and E. ortleppi was found to be highly prevalent in the mithun, with E. ortleppi being reported for the first time. Molecular markers, including the internal transcribed spacer 2 (ITS-2), 28S rDNA and mitochondrial NADH dehydrogenase sub-unit1 (nad1) were used to confirm the identification of the trematode and cestode species.

Genetic characterisation of Taenia multiceps cysts from ruminants in Greece.

This study was designed to genetically characterise the larval stage (coenurus) of Taenia multiceps from ruminants in Greece, utilising DNA regions within the cytochrome c oxidase subunit 1 (partial cox1) and NADH dehydrogenase 1 (pnad1) mitochondrial (mt) genes, respectively. A molecular-phylogenetic approach was used to analyse the pcox1 and pnad1 amplicons derived from genomic DNA samples from individual cysts (n=105) from cattle (n=3), goats (n=5) and sheep (n=97). Results revealed five and six distinct electrophoretic profiles for pcox1 and pnad1, respectively, using single-strand conformation polymorphism. Direct sequencing of selected amplicons representing each of these profiles defined five haplotypes each for pcox1 and pnad1, among all 105 isolates. Phylogenetic analysis of individual sequence data for each locus, including a range of well-defined reference sequences, inferred that all isolates of T. multiceps cysts from ruminants in Greece clustered with previously published sequences from different continents. The present study provides a foundation for future large-scale studies on the epidemiology of T. multiceps in ruminants as well as dogs in Greece.