Use of Pyrosequencing and Denaturing Gradient Gel Electrophoresis to Examine the Effects of Probiotics and Essential Oil Blends on Digestive Microflora in Broilers Under Mixed Eimeria Infection

A protective digestive microflora helps prevent and reduce broiler infection and colonization by enteropathogens. In the current experiment, broilers fed diets supplemented with probiotics and essential oil (EO) blends were infected with a standard mixed Eimeria spp. to determine effects of performance enhancers on ileal and cecal microbial communities (MCs). Eight treatment groups included four controls (uninfected-unmedicated [UU], unmedicated-infected, the antibiotic BMD plus the ionophore Coban as positive control, and the ionophore as negative control), and four treatments (probiotics BC-30 and Calsporin; and EO, Crina Poultry Plus, and Crina PoultryAF). Day-old broilers were raised to 14 days in floor pens on used litter and then were moved to Petersime batteries and inoculated at 15 days with mixed Eimeria spp. Ileal and cecal samples were collected at 14 days and 7 days postinfection. Digesta DNA was subjected to pyrosequencing for sequencing of individual cecal bacteria and denaturing gradient gel electrophoresis (DGGE) for determination of changes in ileal and cecal MC according to percentage similarity coefficient (%SC). Pyrosequencing is very sensitive detecting shifts in individual bacterial sequences, whereas DGGE is able to detect gross shifts in entire MC. These combined techniques offer versatility toward identifying feed additive and mild Eimeria infection modulation of broiler MC. Pyrosequencing detected 147 bacterial species sequences. Additionally, pyrosequencing revealed the presence of relatively low levels of the potential human enteropathogens Campylobacter sp. and four Shigella spp. as well as the potential poultry pathogen Clostridiun perfringens. Pre- and postinfection changes in ileal (56%SC) and cecal (78.5%SC) DGGE profiles resulted from the coccidia infection and with increased broiler age. Probiotics and EO changed MC from those seen in UU ilea and ceca. Results potentially reflect the performance enhancement above expectations in comparison to broilers not given the probiotics or the specific EO blends as feed supplements.

Serological profile of anti-Toxoplasma gondii antibodies in apparently healthy dogs of the city of Botucatu, São Paulo state, Brazil

Toxoplasmosis is a zoonosis caused by Toxoplasma gondii, a coccidian protozoan of worldwide distribution. The seroprevalence in canine population can be an alternative for measuring T. gondii urban spreading. A total of 780 blood samples from dogs were collected, during the yearly anti-rabies campaign, carried out by the Department of Veterinary Hygiene and Public Health, School of Veterinary Medicine and Animal Husbandry (FMVZ), São Paulo State University, UNESP, together with the county health authorities, in August 1999. Using Indirect Fluorescent Antibody Test (IFAT) for detecting antibodies anti-T. gondii in the sera samples, we observed that 258 dogs (33.1%) were positive. The associations between the serological results and the epidemiological variables were studied. Statistically significant differences were not found regarding sex (32.2% male and 34.3% female reactors). Dogs without a defined breed showed seropositivity statistically higher than the pedigreed group. The occurrence of infection was considered higher with age.

Coccidiose em cães e gatos do município de andradina, estado de são paulo, Brasil

Coccidiosis is one of the most common parasitic diseases in dogs and cats in all the world. The aim of this study was to determine the occurrence of this parasitosis in dog and cat population at the Municipality of Andradina in the State of São Paulo, from 2007 to 2009. Fecal samples from 97 cats and 93 dogs were analyzed by using the techniques of flotation in saturated sodium chloride and spontaneous sedimentation. The species were classified according to morphology of the oocysts. Cystoisospora fecal oocyst found in 71.1% (69/97) of the cats, and simple infection by C. rivolta and C. felis occurred respectively in 41.0% (16/39) and 20.5% (8/39) animals, with P ≥ 0.2319. In 39.7%(37/93) of the dogs was found positive for Cystoisospora spp. And the species C. canis identified in the largest proportion (63.9%) with P = 0.0005. From the results, we conclude that dogs and cats had high incidence of infection Cystoisospora, being C. canis and C. rivolta most observed species, respectively.

Sexual transmission of Toxoplasma gondii in sheep

Male sheep of reproductive age were distributed into three groups: GI, a sheep inoculated (oral) with 2.0×105 oocysts of the P strain of Toxoplasma gondii; GII, a sheep infected (subcutaneous) with 1.0×106 tachyzoites of the RH strain of T. gondii; and GIII, a sheep kept as a control (not infected). After the inoculation of the males, 12 breeding ewes, which were not pregnant and which were serologically negative for reproductive diseases (particularly toxoplasmosis), were distributed into three groups, synchronized, and subsequently exposed to natural mating with previously inoculated males. The distribution was as follows: five ewes that underwent natural mating with the GI male, five ewes that were exposed to natural mating with the GII male, and two ewes that were mated with the non-infected male (control). Serum samples of all the ewes were collected on days -30, -14, -7, -1, and 0 (days before natural mating) and on days 1, 3, 5, 7, 11, 14, and weekly until birth; the presence of serum antibodies against T. gondii was assessed by IFAT. Using a bioassay and PCR, T. gondii was isolated from the semen of the infected reproducing sheep before mating. Following natural mating, 5 of the 12 females displayed antibodies specific for T. gondii; of these animals, two of the ewes underwent natural mating with the male inoculated with oocysts (GI) and three with the male infected with tachyzoites (GII). One of the females that displayed antibodies specific to this coccidian and that underwent natural mating with the GII sheep had a macerated fetus on the 70th day following coverage. Using a bioassay after the birth, it was possible to isolate T. gondii from samples of the pool of tissues from the five females that seroconverted after natural mating and from their respective lambs. Using PCR, the DNA of T. gondii was isolated from the pool of tissues from one and two females exposed to natural mating with the reproductive males infected with the oocysts and tachyzoites, respectively. Using this technique, it was also possible to diagnose the presence of the parasite in the pool of tissues from the lambs of one female that underwent natural mating with the male sheep infected with oocysts. These results demonstrated the sexual transmission of T. gondii in the sheep species with consequent vertical transmission to their lambs. © 2013 Elsevier B.V.

Aspectos epidemiológicos da infecção por cryptosporidium em equinos

Inquiries about cryptossporidiosis are essential because its zoonotic potential and pathogenicity in pets and production animals. The horse can be involved in the transmission of the infection by Cryptosporidium and can have an epidemiological role in this disease. The great possibility of adquiring this protozoan, justify the importance of attempting against for the occurrence of this illness. The aim of this article was to demonstrated through literature revision, some epidemiological aspects of the cryptosporidiosis in the equine specie.

Ocorrência de parasitoses gastrointestinais em cordeiros no município de Alambari, São Paulo

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Eimeria species in young and adult sheep raised under intensive and/or semi-intensive systems of a herd from Umuarama city, Parana State, Brazil

The present study aimed to identify Eimeria species in young and adult sheep raised under intensive and / or semi-intensive systems of a herd from Umuarama city, Parana State, Brazil using the traditional diagnostic methods and to correlate the infection level/types of infection in the different age/system in this herd. Fecal samples were collected from the rectum of 210 sheep and were subjected to laboratory analysis to differentiate the species. Furthermore, animals were observed to determine the occurrences of the clinical or subclinical forms of eimeriosis. Out of the 210 collected fecal samples, 147 (70%) were positive for Eimeria oocysts, and 101 (47.86%) belonged to young animals that were raised under intensive and / or semi-intensive farming systems. Oocysts from 9 species of Eimeria parasites were identified in the sheep at the following prevalence rates: E. crandallis, 50.0%; E. parva, 21.6%; E. faurei, 8.1%; E. ahsata, 8.1%; E. intricata, 5.4%; E. granulosa, 2.7%; E. ovinoidalis, 2.0%; E. ovina, 1.3%; and E. bakuensis, 0.6%. There were no differences regarding the more frequent Eimeria species among the different ages of animals or between the different farming management systems. Based on these data, E. crandallis was the most prevalent, followed by E. parva and E. faurei species, regardless of the age. Higher parasitism was diagnosed in the young animals that were raised in a confinement regime, and the disease found in the herd was classified as subclinical. Further studies should be conducted in this herd, to verify if the eimeriosis subclinical can cause damage especially in young animals with a high level of infection.

Efficacy of sulfadoxine plus trimethoprim compared to management measures for the control of Eimeria parasitism in naturally infected and clinically asymptomatic sheep that were maintained in a feedlot

The efficacy of sulfadoxine + trimethoprim in comparison to management measures for the control of Eimeria parasitism was studied in naturally infected sheep that were raised in a feedlot and were clinically asymptomatic for eimeriosis. Weight gain was also evaluated in these animals. The following groups were formed with 15 animals/group: TO!, control animals that received saline solution and maintenance of the same management measures that were performed before the study; T02, animals that received two intramuscular doses of sulfadoxine (20 mg/kg) + trimethoprim (4 mg/kg) with a 14-day interval; T03, sheep that received two intramuscular doses of sulfadoxine (20 mg/kg) + trimethoprim (4 mg/kg) with a 14-day interval plus management measures (wood shaving bedding was changed every Monday, and 30g of ammonium sulfate were applied to the bedding and other facilities were performed every Thursday, 10 mL/20 L of water); and T04, animals that received only the management measures described for the previous group. The highest efficacy rates (arithmetic mean) for the T02 group (sulfadoxine + trimethoprim at days 0 and 14) were 21.04% and 21.98% on the 14th and 28th days after the first treatment (DAFT), respectively. However, the treatment showed efficacy rates below 17% and was totally ineffective from the 70th DAFT to the end of the study. In both the T03 (chemical treatment+ management) and T04 (management only) groups, a significant (P <= 0.05) reduction of oocyst shedding per gram of feces was observed in the animals from the 14th DAFT in comparison to the control group; however, an efficacy rate above 90% was observed from the 28th DAFT. Animals belonging to the T02, T03 and T04 groups presented with alterations in weight gain of 0.57 kg, 4.30 kg and 4.53 kg, respectively, in comparison with the control animals (T01) throughout the 91-day study period. Thus, it is possible to conclude that the two-dose sulfadoxine + trimethoprim treatment, given with a 14-day interval, had little no effect on the oocyst shedding. Moreover, the adopted management measures were enough to cause a significant decrease in the animal parasite loads. (C) 2013 Published by Elsevier B.V.

Pesquisa de Toxoplasma gondii (Nicolle & Manceaux, 1909) em sêmen, testículos e epidídimos de gatos (Felis catus) primoinfectados

Pós-graduação em Medicina Veterinária - FCAV

Validação diagnóstica para a detecção de Cryptosporidium spp. em bovinos

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

A comparative transcriptome analysis reveals expression profiles conserved across three Eimeria spp. of domestic fowl and associated with multiple developmental stages

Coccidiosis of the domestic fowl is a worldwide disease caused by seven species of protozoan parasites of the genus Eimeria. The genome of the model species, Eimeria tenella, presents a complexity of 55-60 MB distributed in 14 chromosomes. Relatively few studies have been undertaken to unravel the complexity of the transcriptome of Eimeria parasites. We report here the generation of more than 45,000 open reading frame expressed sequence tag (ORESTES) cDNA reads of E. tenella, Eimeria maxima and Eimeria acervulina, covering several developmental stages: unsporulated oocysts, sporoblastic oocysts, sporulated oocysts, sporozoites and second generation merozoites. All reads were assembled to constitute gene indices and submitted to a comprehensive functional annotation pipeline. In the case of E. tenella, we also incorporated publicly available ESTs to generate an integrated body of information. Orthology analyses have identified genes conserved across different apicomplexan parasites, as well as genes restricted to the genus Eimeria. Digital expression profiles obtained from ORESTES/EST countings, submitted to clustering analyses, revealed a high conservation pattern across the three Eimeria spp. Distance trees showed that unsporulated and sporoblastic oocysts constitute a distinct clade in all species, with sporulated oocysts forming a more external branch. This latter stage also shows a close relationship with sporozoites, whereas first and second generation merozoites are more closely related to each other than to sporozoites. The profiles were unambiguously associated with the distinct developmental stages and strongly correlated with the order of the stages in the parasite life cycle. Finally, we present The Eimeria Transcript Database (http://www.coccidia.icb.usp.br/eimeriatdb), a website that provides open access to all sequencing data, annotation and comparative analysis. We expect this repository to represent a useful resource to the Eimeria scientific community, helping to define potential candidates for the development of new strategies to control coccidiosis of the domestic fowl. (C) 2011 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.

Controllo dei parassiti nel rettilario del Bioparco di Roma

Poiché le infestazioni parassitarie dei rettili in cattività possono incidere sulla loro salute, è stato condotto uno studio sui parassiti intestinali e gli ectoparassiti su 213 rettili del Bioparco di Roma. In cattività, dove gli animali sono confinati in piccoli spazi, la concentrazione di parassiti può essere più alta. Alcuni di essi sono innocui, ma altri, specialmente in associazione allo stress, possono essere pericolosi, causare malattie e portare anche alla morte se non trattati. Inoltre, i rettili parassitati hanno una vita più breve, sono più suscettibili ad altre malattie, hanno scarsa fertilità, ridotta crescita ed alta mortalità. Tra Gennaio 2012 e Dicembre 2014 sono stati raccolti campioni da 213 rettili che non mostravano alcun segno clinico e che includevano 23 specie di sauri, 20 specie di cheloni, 20 specie di ofidi e 4 specie di alligatori. I campioni sono stati esaminati per la presenza di parassiti intestinali mediante copromicroscopia di tipo qualitativo previa sedimentazione e successiva flottazione, mentre la tecnica di Zihel-Neelsen è stata usata per la ricerca di oocisti di Cryptosporidium . I campioni di feci sottoposti ad esame copromicroscopico qualitativo sono risultati positivi per almeno una tipologia di endoparassiti nel 49% dei casi. Il 35% dei campioni è risultato positivo per ossiuri, il 22% per coccidi, l’1% per strongili ed il 5% per ascaridi. Degli strisci fecali colorati con la tecnica di Zihel-Neelsen, il 25% sono risultati positivi per Cryptosporidium spp. Ossiuri e coccidi sono stati più prevalenti nei sauri rispetto ai cheloni, agli ofidi e agli alligatori e nel 14% è stata osservata la presenza di entrambi. Inoltre, per quanto riguarda gli ectoparassiti, sono state rilevate uova di Myocoptes musculinus nel 24% dei rettili esaminati. Tutti gli strisci risultati positivi per Cryptosporidium spp. sono stati inviati all’Università di Santiago de Compostela per essere sottoposti alla tecnica PCR.

Die Coccidienkrankheit der Kaninchen.

Mode of access: Internet.

Anemia en ovejas de desarrollo en Finca Santa Rosa durante el período Junio –Julio 2014

Con el objetivo de elaborar un plan sanitario que prevenga el síndrome anémico en el hato ovino de la Facultad de Ciencia Animal - UNA. Se determinó la prevalencia de esta patología en los ovinos en desarrollo, Analizando los niveles de infestación ( NI ) de las especies parasitarias y evaluando los valores de hematocrito en relación con la carga gastro - parasitaria. Se tomaron 18 muestras biológicas de heces fecales y sangre, antes de la desparasitación y 20 días después, en el periodo de Junio – Julio 2014, se utilizó la técnica de Sheather, se determinó Hematocrito y Hemoglobina , Los datos s e analizaron a través del estudio epidemiológico descriptivo y analítico utilizando diferentes técnicas de laboratorio y método estadístico de correlación de variables cuantitativas de Pearson . En el primer y segundo muestreo se identificaron los parásitos Haemonchus sp, Cooperia sp, Strongylus sp, Coccidia sp, Bunostomum sp y Moniezia sp . Al primer muestreo la variable prevalencia de síndrome anémico del hato ovino es del 50% y el 100% de los animales analizados presentaron Strongylus sp, Haemonchus sp, Co ccidia sp, Bunostomum sp y Moniezia sp, y el 72,2% presentaron Cooperia sp , presentaron NI Alto 77.7% afectados con Haemonchus sp, 16.6% Strongylus sp, y un 11.1% para las especies de Cooperia sp, Coccidia sp y Moniezia sp. , NI Moderado, Moniezia sp y C occidia sp , un 88.8%, Strongylus sp 83.3%, Bunostomum sp y Haemonchus sp 22.2% y Cooperia sp 11.1%, NI Leve, el 77.7% con Bunostomum sp y el 50% presentaron Cooperia sp. Al segundo muestreo 20 días después del tratamiento, el valor de hematocrito, e staban en los rangos de 22 - 38%. Los niveles de infestación variaron, presentando NI Alta , para Moniezia sp con 22.2%, aumentado su carga parasitaria en relación con el primer muestreo . Se observó la disminución de las cargas parasitarias de las especies Ha emonchus sp con 16.6%, Strongylus sp y Coccidia sp 5.5%.En el NI Moderada , Coccidia sp , mantuvo el 88.8%. Strongylus sp aumentó con 88.8%, Haemonchus sp 83.3%. Las cargas de Moniezia sp , prevalecieron con el 77.7%. Cooperia sp , se presentó con el 50%, B onostomus sp , 27.7%. NI Leve , encontramos el 44.4% de la población con Bunostomum sp y un 5.5% con Cooperia sp y Coccidia sp. De la relación existente entre la presencia de los parásitos y los valores de hematocrito, las combinaciones parasitarias siguient es fueron significativos con P= 0.67, <0.0022 para Cooperia sp y Haemonchus sp , P= 0.65, <0.0036 Cooperia sp con Bunostomum sp , P= 0.62, <0.0060 Cooperia sp con Strongylus sp , P= 0.50, <0.0353 Strongylus sp con Moniezia sp y P= 0.64 <0.0043 para Haemon chus sp con Bunostomum sp. Las parasitosis gastrointestinales son un factor importante de las causas del síndrome anémico, ya que la relación de los valores de hematocritos con la combinación de diferentes especies de parásitos fue significativa y que con la utilización de fármacos antihelmínticos de uso continuo se encontraron cargas parasitarias, con niveles de infestación de alto a moderado .

Evaluación de pollos camperos en producción intensiva y semi-intensiva con suplementación de extracto de quillaja y residuos de hortalizas

En la presente investigación se evaluó: la ganancia diaria de peso, consumo semanal, índice de productividad, mortalidad, conversión alimenticia, costo por kg de carne, pigmentación en tarsos, porcentaje de grasa y el efecto del extracto de quillaja como coccidiostato, bajo dos sistemas de crianza, intensiva y semi-intensiva. El extracto de quillaja fue utilizado al 0,1% de inclusión en el alimento. La investigación se llevó a cabo en la provincia del Azuay, cantón Cuenca, parroquia San Joaquín, sector Balzay Bajo. Se utilizaron 300 pollitos camperos de la estirpe Hubbard variedad redbro S de 1 día de edad. Las aves se distribuyeron de forma aleatoria en un diseño de bloques al azar con 3 tratamientos, cada uno con 5 repeticiones y con 20 pollitos por unidad experimental. Los tratamientos fueron: T1: testigo, aves alojadas en sistema intensivo; T2: aves alojadas en sistema intensivo, más una dieta modificada que consistía en la adición de extracto de quillaja al 0,1%; T3: sistema semi-intensivo con la misma dieta del T2, las aves de este tratamiento a partir del día 28 de edad tuvieron acceso a un área verde delimitada, la cual poseía una mezcla forrajera de raigrás-alfalfa y además se adicionaron a su alimentación residuos de hortalizas propias de la zona. La investigación duró 56 días, no se evidenciaron diferencias significativas en ganancia diaria de peso, índice de productividad, índice de conversión, costo por kg de carne y porcentaje de grasa (p>0,05), mientras que consumo semanal y mortalidad, mostraron diferencias significativas (p<0,05). En las demás variables se evidenció mejor intensidad de pigmentación en T3 (p<0,05), mientras que en la infestación por coccidios no se observó diferencia entre tratamientos (p>0,05), lo que indica que la aplicación de extracto de quillaja tuvo un efecto similar al programa anticoccidial utilizado en T1