Screening Mediterranean shrubs selected by browsing goats against gastrointestinal strongyles

Ethanolic extracts from the main species selected by browsing Charnequeira goats on Mediterranean vegetation were assayed for a rapid screening of their anthelmintic potential against gastrointestinal (GI) strongyles. Shrubs selected included Olea europeae var. sylvestris (OS), Quercus coccifera (QC), Pistacia lentiscus, Rhamnus alaternus and Rhamnus lycioides (RL). Forage species included Cichorium intybus (CI) and Hedysarum coronarium (HC). Extract concentrations were chosen taking as reference the fecal total phenolics content (TP) from those goats after a five-month browsing period (8.4 mg GAE/g DM). A quantitative coproculture method was adapted using faeces obtained from naturally infected goats, not dewormed or having access to browse (reference group); larvae culture was performed in triplicate for three different TP concentrations (25, 10 and 5 mg GAE/g DM) against a distilled water control, for a 7-day incubation period at 27 °C. The number of larvae developed was counted differentiating total and live larvae per gram of faeces (LDPG). The efficacy of each extract and concentration, as percentage of reduction in LDPG against control, was determined. Data was log transformed and submitted to analysis of variance. Means were compared by Tukey test at 5% significance level.A reduction in LDPG was observed in all extracts being significant (P<0.05) for the highest concentration. The extracts’ efficacy over exogenous forms showed not only a reduction of total LDPG but, for some, also a high level of larvae mortality, resulting in reduction of live infective larvae (L3). These results indicate that some extracts, besides their ovicidal activity, have also larvicidal potential. Extracts efficacy for total LDPG was higher for QC (82%), OS (80%), HC and RL (76%) and CI (69%). Reduction of live L3 was higher for OS (90%) and HC (80%). Results obtained on these extracts potential in reducing exogenous forms, namely L3 forms of GI strongyles, is an important aspect in the control of these parasitic populations, as it results in lower pasture contamination and consequently a lower rate of reinfection of the animals. Funding obtained from project VegMedCabras – ALT20-03-0145-FEDER-000009

Screening Mediterranean shrubs selected by browsing goats against gastrointestinal strongyles

Ethanolic extracts from the main species selected by browsing Charnequeira goats on Mediterranean vegetation were assayed for a rapid screening of their anthelmintic potential against gastrointestinal (GI) strongyles. Shrubs selected included Olea europeae var. sylvestris (OS), Quercus coccifera (QC), Pistacia lentiscus, Rhamnus alaternus and Rhamnus lycioides (RL). Forage species included Cichorium intybus (CI) and Hedysarum coronarium (HC). Extract concentrations were chosen taking as reference the fecal total phenolics content (TP) from those goats after a five-month browsing period (8.4 mg GAE/g DM). A quantitative coproculture method was adapted using faeces obtained from naturally infected goats, not dewormed or having access to browse (reference group); larvae culture was performed in triplicate for three different TP concentrations (25, 10 and 5 mg GAE/g DM) against a distilled water control, for a 7-day incubation period at 27 °C. The number of larvae developed was counted differentiating total and live larvae per gram of faeces (LDPG). The efficacy of each extract and concentration, as percentage of reduction in LDPG against control, was determined. Data was log transformed and submitted to analysis of variance. Means were compared by Tukey test at 5% significance level.A reduction in LDPG was observed in all extracts being significant (P<0.05) for the highest concentration. The extracts’ efficacy over exogenous forms showed not only a reduction of total LDPG but, for some, also a high level of larvae mortality, resulting in reduction of live infective larvae (L3). These results indicate that some extracts, besides their ovicidal activity, have also larvicidal potential. Extracts efficacy for total LDPG was higher for QC (82%), OS (80%), HC and RL (76%) and CI (69%). Reduction of live L3 was higher for OS (90%) and HC (80%). Results obtained on these extracts potential in reducing exogenous forms, namely L3 forms of GI strongyles, is an important aspect in the control of these parasitic populations, as it results in lower pasture contamination and consequently a lower rate of reinfection of the animals. Funding obtained from project VegMedCabras – ALT20-03-0145-FEDER-00000

Mudança de paradigma na abordagem aos parasitas gastrointestinais em pequenos ruminantes – controlo integrado com tratamento seletivo

O crescente desenvolvimento de resistências aos anti-helmínticos, com particular importância em pequenos ruminantes, tem ameaçado a sustentabilidade da produção no sector. Este desafio no controlo é exacerbado pela escassez de moléculas alternativas e pela crescente consciencialização da contaminação ambiental aquando da excreção dos compostos anti-helmínticos administrados. Neste trabalho, combinaram-se os dados preliminares recolhidos ao longo de dois projectos independentes (VegMedCabras e CistusRumen financiados pelo Fundo Europeu para o Desenvolvimento regional, Alentejo 2020), ainda em curso. Enquanto o primeiro visa promover o pastoreio da vegetação arbustiva mediterrânica de forma ecológica e sustentável por caprinos, o segundo tem por objectivo uma utilização sustentável da Esteva (Cistus ladanifer L.), em pequenos ruminantes, como ferramenta no aumento da competitividade e na redução do impacte ambiental. Em ambos, a infecção parasitária tem vindo a ser avaliada individualmente através da determinação do número de ovos eliminados por grama de fezes (OPG) e aplicada a técnica qualitativa de flutuação por Willis em todas as amostras, individualmente. Os ensaios experimentais realizados no âmbito do projecto CistusRumen incidiram sobre borregos (36 animais) e cabras adultas (18 animais), sujeitos a dietas suplementadas com diferentes concentrações de taninos ou esteva. Entre os resultados obtidos, os achados dos exames qualitativos foram os que revelaram o maior potencial dos taninos. Durante as análises coprológicas dos animais pertencentes a grupos com dietas suplementadas registou-se um fenómeno de rebentamento ou alteração da morfologia dos ovos de estrongilídeos. Os resultados preliminares recolhidos nos ensaios experimentais do CistusRumen complementam os objectivos do VegMedCabras, uma vez que a incorporação de vegetação arbustiva mediterrânica, como a Esteva, cujos taninos auxiliam a absorção proteica ao nível do intestino delgado, contribuem para uma maior capacidade de resposta do hospedeiro às infecções parasitárias. Um aumento na capacidade de resposta associada ao aparente efeito da suplementação da dieta sobre as formas exógenas diminui a necessidade do controlo pelo tratamento, reservando-o para casos específicos com tratamento selectivo do efectivo diminuindo assim a pressão de selecção sobre as resistências e diminuindo a contaminação ambiental

Infestações de carraças (Acari:Ixodidae) em populações de bovinos na bacia do Rio Geba, Guiné- Bissau

As carraças são ectoparasitas de grande importância veterinária, em regiões tropicais, pela sua acção espoliadora de sangue do hospedeiro e pela sua capacidade de agir como vectores de agentes patogénicos. De modo a identificar as espécies de carraças em populações de bovinos, na bacia do Rio Geba (regiões de Bafatá e Gabú), na Guiné-Bissau, foram seleccionados 149 bovinos, no total, para inspecção quanto à presença de ectoparasitas que levou à colheita de 337 carraças. Em cada manada foram inspeccionados visualmente entre 5 a 10 animais e os produtores responderam a um questionário para avaliação do tipo de maneio e de controlo parasitário efectuado. O estudo abrange 19 produtores de 18 tabancas, 11 dos quais em Abril de 2010 (estação seca) e outros 8 em Setembro de 2012 (estação húmida). Foram identificados 3 géneros de carraças – Amblyomma (51,3%), Hyalomma (0,9%) e Rhipicephalus (47,8%) – que incluem 5 espécies (Ambyomma variegatum, Hyalomma truncatum, Rhipicephalus (Boophilus) annulatus, Rhipicephalus (Boophilus) geigyi e Rhipicephalus sulcatus). As espécies do género Amblyomma foram, claramente, predominantes durante a estação húmida (88.9%) enquanto as espécies pertencentes ao género Rhipicephalus foram as mais abundantes na estação seca (75,7%). Apesar dos produtores reconhecerem que as doenças transmitidas por carraças constituem um problema na região continuam a exercer medidas de controlo tradicionais. Independentemente das diferenças registadas entre manadas e entre as estações seca e húmida, as espécies encontradas estão em concordância com os dados anteriores registados na regiã

CHEYLETIELLA PARVOSITIVORAX: PRIMEIRO REGISTO DE UMA INFESTAÇÃO EM COELHOS EM MOÇAMBIQUE

Introdução: Os ácaros do género Cheyletiella pertencem à Ordem Prostigmata, Família Cheyletidae. Três espécies são descritas nos animais domésticos nomeadamente, C.yasguri em cães, C. blakei em gatos e C. parasitivorax em coelhos (Taylor et al., 2007). Estas têm distribuição mundial e são consideradas zoonóticas, podendo infectar humanos quando em contacto directo com algum portador (Soulsby, 1982). Os ácaros deste género vivem permanentemente nos pelos, aproximando-se da pele apenas para se alimentarem. As fêmeas durante a ovopostura colam os aos pêlos. Destes eclodem as larvas, que passam por dois estádios ninfais antes de atingirem a fase adulta. O ciclo completo dura aproximadamente duas semanas Objectivos: O presente poster tem como objectivos reportar a ocorrência da Cheyletiella parasitivorax em Moçambique, contribuir com elementos que permitam o correcto diagnóstico deste ácaro e alertar para a importância zoonótica deste parasita. Materiais e Métodos: Foram colhidas amostras de pelo e raspagens cutâneas de coelhos com sinais de alopécia, descamação e prurido. As amostras foram processadas pelo método de clarificação com lactofenol de Amman Resultados: Foram observados ácaros que apresentavam como principais características morfológicas: forma oval, tamanho aproximado de 400 m, patas grandes, palpos com artículos terminais em forma de garra. As características morfológicas e morfométricas conduziram à sua classificação como Cheyletiella parasitivorax. Discussão: É a primeira vez que um caso de um coelho parasitado com C. parasitovorax é reportado em Moçambique. Por este facto, é necessário chamar a atenção de todos profissionais de veterinária para o correcto diagnóstico e consequentemente tratamento eficaz desta zoonose

Plano de Estudo Epidemiológico do Porco Alentejano e Cruzado

Com o objectivo de estudar a patologia associada à produção de porcos de raça Alentejana, a Divisão de Saúde Animal dos Laboratórios Pfizer, a empresa Eservet e a Universidade de Évora, através do Hospital Veterinário e do Departamento de Medicina Veterinária estabeleceram um protocolo de colaboração para levar a efeito o Plano de Estudo Epidemiológico do Porco Alentejano e Cruzado – PEEPAC. Os produtores e médicos veterinários assistentes de explorações de suínos de raça Alentejana e/ ou resultantes de cruzamentos com raça Alentejana têm beneficiado, de acordo com o protocolo, de um serviço de apoio ao diagnóstico constituído por um conjunto de análises laboratoriais. As amostras enviadas ao laboratório para análise, consistindo em cadáveres ou vísceras de um ou mais animais, provenientes de diferentes explorações, foram submetidas a exames anatomopatológicos, bacteriológicos e parasitológicos. A pesquisa de vírus foi apenas efectuada quando especificamente solicitada pelo médico veterinário assistente na respectiva exploração. Serão apresentados e discutidos os resultados dos exames necrópsicos e exames histopatológicos; os resultados da pesquisa de bactérias aeróbias, de bactérias anaeróbias e respectivos testes de sensibilidade a agentes antimicrobianos; os resultados dos exames coprológicos, bem como das análises macro e microscópicas para pesquisa de ecto e endoparasitas

Evaluation of Cistus ladanifer L. effect on gastrointestinal parasites in lambs

Evaluation of Cistus ladanifer L. effect on gastrointestinal parasites in lambs Inês Delgadoa, Sara Tudela Zúquete, David Soldado, Olinda Guerreiro, Eliana Jerónimo, Ludovina Neto Padre Cistus ladanifer L. (Cistaceae) is a bountiful available shrub at the western Mediterranean region, including the south of France, Spain, Portugal and the north of Morocco. It is commonly observed in uncultivated fields. This perennial shrub contains low protein levels and high levels of phenolic compounds, such as condensed tannins (CT). It has a low organic matter digestibility, which, enhanced by its anti-nutritional components results in a poor nutritional value. However, either by natural grazing or by encouraged consumption, the incorporation of Cistus ladanifer L. in small ruminants’ diet may simultaneously act as an important feeding complement as it can transform this shrub into a high-level end-use product. With this objective, our group wanted to determine if its composition could have an influence on gastrointestinal parasites. Gastrointestinal nematode and coccidia pose as a highly significant economic burden for small ruminant production systems. Phenolic compounds, such as tannins are associated with a decrease in gastrointestinal parasites, namely, nematodes. In this study, we evaluated the effect of three levels of Cistus ladanifer CT (0, 1.25 and 2.5%) and two ways of CT supply (C. ladanifer aerial parts and C. ladanifer CT extract) on on gastrointestinal parasites in lambs. Thirty-six crossbred Merino Branco x Romane ram lambs of approximately 60 days of age were randomly assigned to individual pens (6 per dietary treatment). A total of 4 stool collections were sampled from each animal – before dietary treatment application (day 0), at the end of the adaptation period of diets (day 7), and at days 24 and 42 of the trial. Coprological techniques included nematode egg (Epg) and coccidian count by the concentration technique McMaster and all samples were also evaluated by direct microscopic observation by the Willis fluctuation method. Data was analyzed using a generalized linear mixed model procedure that identified a significant interaction of treatment groups over time regarding the strongylid Epg count (Chi2 = 85.9, p ≤ 0.001), through the Penalized Quasi-Likelihood parameter estimation method. Pairwise contrast tests, with the Holm adjustment method, showed that both ways of C. ladanifer CT supply at level of 1.25% of CT (C. ladanifer aerial parts - Chi2 = 21.7, p = 0.0006; and C. ladanifer CT extract - Chi2 = 21.4, p = 0.0006) resulted in significantly lower Epg counts of strongylids over time when compared to treatments without CT supply. This work is funded by Alentejo2020 program through the FEDER under the project CistusRumen (ALT20-030145-FEDER-000023) and by National Funds through FCT under the Project UID/AGR/00115/2019

Tick-borne diseases in asymptomatic cattle from São Miguel Island, Azores – A clinical perspective

Tick-borne diseases in asymptomatic cattle from São Miguel Island, Azores – A clinical perspective Introduction/Background: The Azores archipelago is one of the regions with the highest cattle production, in Portugal. Outbreaks of tick-borne (TB) diseases have been reported by local field veterinarians and different tick species are described in São Miguel. At this study, we decided to investigate the presence of some of these agents in asymptomatic cattle, in S. Miguel. Materials & methods: In October 2019, 10 blood samples from each of 10 farms were randomly selected. Species belonging to the Babesia/Theileria and Anaplasma/ Ehrlichia genus were tested by PCR screening. Results: There were 45 positive samples later confirmed as species belonging to the Theileria orientalis group (45/100, 45%), in 9 farms (9/10, 90%) and it was also possible to identify Anaplasma bovis, in 2 animals, at two farms (2/10, 20%). Discussion: As far as we were able to verify, this is the first reference to the presence of these agents in S. Miguel. However, although asymptomatic, the repercussions associated with these infections are probably being underestimated or confused with other health problems. Since ixodids were not found in cows, other factors may be contributing to the high percentage of positive samples found, such as poor management practices. This is clearly an interesting subject for further investigation, as a better understanding of the life cycle of these parasites will contribute to their control

Evaluating natural alternatives as hypothetical nutraceuticals against gastrointestinal parasites in lactating Goats

Evaluating natural alternatives as hypothetical nutraceuticals against gastrointestinal parasites in lactating Goats Sara Tudela Zúquetea, Inês Delgado, David Soldado, Liliana Cachucho, Letícia Fialho, Olinda Guerreiro, Eliana Jerónimo, Ludovina Neto Padre E-mail: [email protected] Gastrointestinal parasite burden on goats translates into high production losses due to inadequate growth and poor milk production. These parasites are also responsible for higher mortality rates when facing high infestations levels or, more frequently, in kids. During the last decades, effective anti-helminthic compounds controlled such situations. However, the approach to such problem shifted and marked animal production systems from the eighties onwards. Drugs used for treatment of severe situations started to be administered as prophylaxis at frequent time intervals. Initially, such switch had an enormous success as it allowed an unforeseen growth and intensification of animal production at a relative low cost. Later, the onset of the first anti-helminthic resistance was bypassed by new molecule research and synthesis. Notwithstanding, presently, resistance to the most recent anti-helminthics, macrocyclic lactones and monepantel, are reported at distinct regions of the globe, even though some countries have not yet approved the latter, as the US. Clinicians are now facing a new threat as we are on the verge to lose the molecules we use to treat. Furthermore, resistance also poses as a public health threat not only for the risks implied specially for soiltransmitted helminthes subjected to the pharmaceutical compounds excreted by animal feces, but also by the environmental contamination only recently being consciously addressed. In order to surpass such threats, new natural alternatives must be further investigated. At the present work, we evaluated the effect of two distinct condensed tannin (CT) plant extracts – Cistus ladanifer L. (Rockrose, Esteva in Portuguese) and Schinopsis lorentzii (Quebracho) on gastrointestinal parasites in lactating goats. Eighteen Serpentina goats were individually housed and randomly assigned to one of three diets (control – without CT; Ci – with C. ladanifer extract; Sc – with S. lorentzii extract). In both diets with CT extracts, 20 g/head of CT were supplied daily. Individual stool collections were sampled before dietary treatment application (day 0), at the end of the adaptation period of diets (day 14), and at days 21, 28 and 35 of the trial. Coprological analysis included concentration McMaster technique for egg count (Epg) and Willis fluctuation assay for microscopic observation. Data was analyzed using a generalized linear mixed model procedure that identified a significant interaction of treatment groups over time regarding the strongylid Epg count (Chi2 = 1355.6, p ≤ 0.001), through the Gauss-Hermite quadrature parameter estimation method. Pairwise contrast tests, with the Holm adjustment method, showed that both Ci (Chi2 = 148.1, p < 0.0001) and Sc (Chi2 = 962.3, p < 0.0001) treatments resulted in significantly lower Epg counts of strongylids over time when compared to the control group. The Sc group also presented a significantly lower Epg counts of strongylids over time when compared to the Ci group (Chi2 = 1054.8, p < 0.0001). This work is funded by Alentejo2020 program through the FEDER under the project CistusRumen (ALT20-03-0145FEDER-000023) and by National Funds through FCT under the Project UID/AGR/00115/2019

Mediterranean Shrubs: Nutraceuticals for sustainable goat production

Mediterranean Shrubs: Nutraceuticals for sustainable goat production Costa, C.ab, Almeida, S.M.c, Cavaco-Gonçalves, S.a, Barbas, J.P.a, Ribeiro, J.M.B.F.a, Belo, C.C.a, Padre, L.b, Belo, A.T,a aINIAV, Santarém – National Institute of Agrarian and Veterinarian Research, Quinta da Fonte Boa, 2005-048 Vale de Santarém bICAAM – Department of Veterinary Medicine, Évora University, Pólo da Mitra, 7006-554 Évora cUTAD – Trás-os Montes and Alto Douro University, Quinta de Prados, 5000-801 Vila Real E-mail: [email protected] Bioactive natural products have become an important research topic and development area in different fields of study, owing to their numerous potentialities. In animal production, the anthelminthic, antioxidant and antimicrobial properties of natural vegetation can improve the animals’ productive efficiency and wellbeing, and product quality (meat and milk). It is known that goats have a very selective feeding behavior and a higher sensitivity to gastrointestinal strongyles (GIS) infections compared to other ruminants, and that certain plants have beneficial effects in animal health through the synergy between their nutritive value and bioactive compounds. Thus, the project “VegMedCabras” intends to characterize and explore the nutraceutical value of mediterranean shrubs for sustainable goat production, by controlling parasitic infection as well as preventing environmental contamination and residues in the final product from the continuous use of synthetic anthelmintics [1,2,3]. Charnequeira breed goats were observed during browsing/grazing of Mediterranean vegetation, from February to July, in order to document their shrub species preferences and evaluate the nutraceutical value of the final diets. All selected plants were analyzed for nutritive value (protein and fiber content, digestibility), as well as bioactive compounds content such as total phenolic compounds (TPC), total tannins (TT), condensed tannins, flavonoids and saponins. Antioxidant activity was also evaluated. Each goat was individually evaluated for natural GIS infection, by egg elimination levels, and health status, by metabolic blood parameters. A significant drop in EPG (eggs per gram of feces) elimination (± 65%) was observed by the end of the study period. As soon as goats started browsing the Mediterranean vegetation, EPG counts started to decrease as the proportion of shrubs rich in TPC increased in the diet. Species with the highest contribution were Pistacia lentiscus and Quercus coccifera, which presented an average TPC and TT of 163,6 and 134,2 mg GAE (gallic acid equivalent)/g MS vs. 138,4 and 130,2 mg GAE/g MS, respectively. Nutritive value of the diets selected by goats was estimated in order to determine the adequacy to their physiological requirements. With the increased resistance of GIS to synthetic anthelmintics and social demand for quality products, it is important to establish multidisciplinary options such as the use of nutraceutical shrubs, a safer, ecological and inexpensive solution. Keywords: nutraceuticals; Mediteranean shrubs; bioactive compounds; anthelmintics; goats References: [1]. Jasmim RS (2018) Int. J. Botany 14: 24-29. [2]. Saric T, Rogosic J, Zupan I, Beck R, Bosnic S, Sikic Z, Skobic D, Tkalcic S (2015) Small Rum. Res. 123: 179-182. [3]. Hoste H, Torres-Acosta JFJ, Sandoval-Castro CA, Muller-Harvey I, Sotiraki S, Louvandini H, Thamsborg SM, Terrill TH (2015) Vet. Parasitology 212: 5-17. Acknowledgements: VegMedCabras – Mediterranean shrubs: natural anthelmintics in the diet selected by grazing goats (ALT-03-0145-FEDER-000009) co-financed by ALENTEJO2020 program-European Regional Development Fund