Cryptosporidiosis and microsporidiosis as causes of diarrhea in kidney and/or pancreas transplant recipients.

INTRODUCTION: Gastrointestinal disorders in solid organ recipients may have various origins including cryptosporidiosis and microsporidiosis. The prevalence of these infections is poorly known in solid organ transplant (SOT) patients in industrialized countries. METHODS: We prospectively assessed the infectious causes of diarrhea in SOT patients. Secondary objectives were to gain further insight into the main characteristics of cryptosporidiosis, and to assess risk factors for this infection. All adult kidney and/or pancreas recipients presenting with diarrhea and admitted to our facility between May 1, 2014 and June 30, 2015 were enrolled. A stool sample was analyzed using a standardized protocol including bacteriological, virological, and parasitological investigations. Data related to clinical symptoms, immunosuppression, and environmental potential risk factors were collected through a self-administered questionnaire and computerized medical records. RESULTS: Out of 73 enrolled patients, 36 had infectious diarrhea (49.3%). Viruses ranked first (17/36), followed by parasites and fungi (11/17). Cryptosporidiosis was the most common parasitic disease (n=6 patients). We observed four microsporidiosis cases. The estimated prevalence of cryptosporidiosis and microsporidiosis in this cohort was 3.7 and 2.40/00, respectively. No significant risk factor for cryptosporidiosis or microsporidiosis, neither environmental nor immunological, could be evidenced. CONCLUSION: Both cryptosporidiosis and microsporidiosis represent a significant cause of diarrhea in kidney transplant recipients.

[The significance of cryptosporidiosis for the health of calves in Switzerland]. / Die Bedeutung der Cryptosporidiose für die Kälbergesundheit in der Schweiz.

INTRODUCTION: Diarrhea in calves is one of the most important cattle diseases in Switzerland. The diagnosis and treatment of calf diarrhea represent a major challenge. Single-celled Cryptosporidium parasites are the most prevalent causative agents of calf diarrhea besides rotavirus in the first weeks of life, and are responsible for about 50% of diarrheal cases. Cryptosporidium parvum has been described as a cause of diarrhea in one to three weeks old calves since the 1970s. Oral ingestion of persistent environmental oocysts results in severe diarrhea lasting four to six days and shedding of large numbers of infectious oocysts. A tiny amount of 10 oocysts is already sufficient to cause disease. Detailed knowledge about the epidemiology and virulence of the different C. parvum strains is still lacking. In addition, current diagnostic tests cannot reliably distinguish between non-pathogenic (e.g. C. bovis) and pathogenic Cryptosporidium species. Until now, no effective therapeutic drug or vaccine against calf cryptosporidiosis has been found. Water-borne epidemics and the zoonotic potential of Cryptosporidium in immunodeficient patients are of great medical importance. The increasing number of cryptosporidiosis cases associated with high infant mortality in less industrialized and impoverished regions (including South-East Asia and sub-Saharan Africa) has intensified the research in recent years. The recent discoveries of new therapeutics against C. parvum may benefit calf medicine in the near future. This review article reports on these new developments, highlights calf cryptosporidiosis in Switzerland and draws attention to a new research project.

INTRODUCTION: La diarrhée chez les veaux est l'une des maladies du bétail les plus courantes en Suisse. Le diagnostic de la cause et le traitement de la diarrhée des veaux représentent un défi majeur. En Suisse, les cryptosporidies sont, avec les rotavirus, l'agent causal le plus fréquent de diarrhée du veau dans les premières semaines et elles sont responsables d'environ 50% des cas. Le parasite unicellulaire Cryptosporidium parvum a été décrit depuis les années 1970 comme un agent de diarrhée chez les veaux d'une à trois semaines. Après ingestion orale d'oocystes persistants dans l'environnement, il se produit après quelques jours une diarrhée sévère de quatre à six jours avec excrétion massive d'oocystes déjà infectieux. Même quelques oocystes persistants dans l'environnement peuvent être pathogènes. Du point de vue épidémiologique, il existe encore de grandes lacunes dans la connaissance de la variabilité suspectée dans la virulence de diverses souches de C. parvum. En outre, des espèces non pathogènes (entre autres Cryptosporidium bovis) peuvent être présentes chez les veaux, qui ne se distinguent pas de C. parvum avec les tests diagnostiques actuels. Jusqu'à présent, aucun médicament efficace sur le plan thérapeutique et aucun vaccin contre la cryptosporidiose du veau n'ont été trouvés. En médecine humaine, les épidémies transmises par l'eau (en particulier aux États-Unis) et l'importance zoonotique des cryptosporidies comme pathogènes opportunistes chez les personnes immunodéficientes jouent un rôle de premier plan. La forte morbidité de la cryptosporidiose associée à une forte mortalité infantile dans les régions les moins industrialisées et les plus pauvres (entre autres en Asie du Sud-Est et en Afrique subsaharienne) ont relancé la recherche sur ces parasites au cours des dernières années. En particulier, la découverte de nouveaux médicaments contre C. parvum est susceptible de bénéficier à la médecine du veau dans un proche avenir. Cet article de synthèse fait le point sur ces nouveaux développements mais surtout sur la cryptosporidiose du veau en Suisse et attire l'attention sur un nouveau projet de recherche.

Cryptosporidium occurrence in ruminants from the North Pioneer mesoregion of Paraná, Brazil / Ocorrência de Cryptosporidium em ruminantes da mesorregião norte pioneiro do Estado do Paraná, Brasil

Abstract The aim of this study was to investigate the occurrence of Cryptosporidium in cattle and sheep from the North Pioneer mesoregion of the state of Paraná. For this, 317 stool samples were collected from cattle and sheep on 16 properties in six municipalities in the North Pioneer mesoregion of Paraná. For detection of Cryptosporidium species, molecular analysis was performed using nested-PCR techniques targeting the 18S rRNA gene. Of the 37 beef cows and 115 calves analyzed, four (10.8%) and 14 (12.2%), respectively, were positive for Cryptosporidium. Of the 12 cows and 52 calves, one (8.3%) and 14 (26.9%), respectively, were positive for Cryptosporidium; and of the 42 ewes and 59 lambs, six (14.3%) and 12 (20.3%), respectively were positive for Cryptosporidium. Cattle (15.3%) and sheep (17.8%) were both susceptible to infection. All the properties of the municipalities of Assaí, Ibaiti and, Leópolis presented infected animals. The study showed that Cryptosporidium occurs in most municipalities assessed, that dairy calves had a higher risk (Odds Ratio=2,66, p-value=0,018) for infection than beef calves, and that sheep are just as susceptible to infection as are cattle, and that further Cryptosporidium studies are developed.

Resumo O objetivo deste estudo foi investigar a ocorrência de Cryptosporidium em bovinos e ovinos da mesorregião norte pioneiro do Estado do Paraná. Para tanto, 317 amostras de fezes destes ruminantes foram colhidas de 16 propriedades de seis municípios do Norte Pioneiro do Paraná. Para detecção de Cryptosporidium spp foi realizada análise molecular pela Técnica de nested-PCR direcionada ao gene 18S rRNA. Das 37 vacas de corte e 115 bezerros de corte analisados, quatro (10,8%) e 14 (12,2%) foram respectivamente positivos para Cryptosporidium . Das 12 vacas e 52 bezerros de leite, um (8,3%) e 14 (26,9%) foram positivos para Cryptosporidium e das 42 ovelhas e 59 cordeiros avaliados, seis (14,3%) e 12 (20,3%) amostras estavam positivas para Cryptosporidium, respectivamente. Bovinos (15,3%) e ovinos (17,8%) foram igualmente suscetíveis à infecção. Todas as propriedades dos municípios de Assaí, Ibaiti e Leópolis apresentaram animais infectados. Este estudo demonstrou que Cryptosporidium ocorre na maioria dos municípios avaliados, sendo que os bezerros de leite apresentam maior risco (Razão de chances=2,66, p-value=0,018) à infecção que os bezerros de corte e que os ovinos são tão suscetíveis à infecção quanto os bovinos e por isso, estudos nesta espécie animal devem ser mais desenvolvidos.

[Clinical epidemiology of cryptosporidiosis in calves]. / Klinische Epidemiologie der Kälber- Cryptosporidiose.

INTRODUCTION: The aim of this study was to determine whether there is an association between Cryptosporidium infections in calves and immunological factors, as well as farm-related factors or the application of the anti-cryptosporidiosis drug Halofuginone. From January to June 2010, 63 cow-calf-pairs from 20 different farms near Zürich, Switzerland have been investigated. Each cowcalf- pair was visited three times within the first 6 weeks of life to collect data of the farm and animals, as well as blood, faecal, colostral and milk samples. An ELISA using sporozoite antigen was developed for the specific detection of anti-Cryptosporidium-IgG in blood- and colostral serum. The IgG concentration in the bloodand colostral serum was determined using radial immuno diffusion test (RID). White blood cell isolation and differential blood cell counts and California Mastitis Test were performed. Bacteriological studies on quarter-milk-samples were carried out. Cryptosporidium oocysts were diagnosed with the modified Ziehl-Neelsen staining, other protozoa with the SAFC method and Eimeria oocysts and helminth eggs were diagnosed with the combined sedimentation/floatation test. ELISAs were performed for the detection of rota- and coronavirus, E. coli F5 and Cryptosporidium spp. in bovine feces (bio-X Diagnostics®, Belgium). The highest prevalence of Cryptosporidium oocysts was 54.0% and found 7 to 20 days post natum, whereas 47.1% were suffering from diarrhea. The transfer of total IgG with the colostrum and the humoral immunity of the calf could not prevent any infection with Cryptosporidium, but the severity of the diarrhea symptoms decreased with increasing total IgG concentrations. Calves housed in open sheds showed significantly more often diarrhea, i. e. they shed more Cryptosporidium oocysts during the first 4 days and 7 to 20 days post natum, respectively. Halofuginone (Halocur®) is approved for prophylaxis against cryptosporidiosis, but it showed no effect on the excretion of Cryptosporidium oocysts in the present study.

INTRODUCTION: Le but de la présente étude était d'étudier s'il existe un rapport entre l'apparition de cryptosporidies et des facteurs immunologiques, des facteurs liés à l'exploitation ainsi qu'à l'usage d'halofuginone. De janvier à juin 2010, on a examiné 63 paires mère-veau provenant de 20 exploitations du canton de Zürich. Au cours de 6 semaines on a effectué, à des moments choisis, trois visites. A ces occasions, des données relatives à l'exploitation ainsi que des échantillons de sang, de selles, de colostrum respectivement de lait ont été collectés. On a développé un test ELISA avec des antigènes de sporozoïtes pour mettre en évidence la présence IgG anti-cryptosporidies dans le sang et dans le colostrum. La concentration en IgG dans le colostrum et dans le sérum a été mesurée avec un test d'immunodiffusion radiale (RID). En outre on a réalisé une image sanguine différentielle des vaches et des veaux et effectué un test de Schalm chez les vaches. Un examen bactériologique a été réalisé sur un échantillon provenant des quatre quartiers. Les oocystes de cryptosporidies ont été mis en évidence au moyen d'une coloration de Ziehlk-Neelsen modifiée, les autres protozoaires ont été mis en évidence par la méthode SAFC et les oeufs d'helminthes ainsi que les oocystes d'Eimeria par un processus de sédimentation-flottation combiné. Un test ELISA a été utilisé pour les rota- et les coronavirus, les E. coli F5 et Cryptosporidium spp. dans les selles des bovins (Bio-X Diagnostics®, Belgique). La prévalence d'infections par des cryptosporidies était maximale entre le 7ème et le 20ème jour de vie des veaux (50.4%), 47.1% de ces veaux souffrant de diarrhée. Les stabulations libres augmentaient de façon significative le risque de diarrhée et d'excrétion de cryptosporidies entre le 1er et le 4ème jour respectivement entre le 7ème et le 20ème jour. La transmission d'IgG et l'immunité humorale des veaux n'empêchaient pas l'infection par des cryptosporidies mais la gravité de la diarrhée diminuait avec l'augmentation de la concentration des IgG totales. L'halofuginone, substance enregistrée pour la prophylaxie de la cryptosporidiose, n'a pas montré, dans cette étude, d'efficacité pour empêcher l'excrétion d'oocystes de cryptosporidies.

Molecular characterisation of Cryptosporidium spp. in lambs in the South Central region of the State of São Paulo / Caracterização molecular de Cryptosporidium spp. em cordeiros na região centro sul do Estado de São Paulo

Considering the proximity of sheep farmers to animals that are possibly diseased or releasing fecal oocysts into the environment and the marked pathogenicity in lambs, the aim of this study was to determine the occurrence and to molecularly characterize the infection by Cryptosporidium spp. in lambs in the South Central region of the state of São Paulo, Brazil. A total of 193 fecal samples were collected from sheep of several breeds, males and females, aged up to one year. Polymerase chain reaction (nested-PCR) was used to amplify DNA fragments from the subunit 18S rRNA gene and indicated 15% positivity; sequencing of amplified fragments was possible for 19 samples. Analysis of the obtained sequences showed that the identified species were Cryptosporidium xiaoi for 15 samples, constituting thus the first molecular characterization study of this Cryptosporidium species in Brazil. Cryptosporidium ubiquitum was identified for three samples and Cryptosporidium meleagridis for one sample; the latter two are considered zoonotic species.(AU)

Devido à proximidade de criadores de ovinos com animais possivelmente doentes e/ou eliminando oocistos fecais no ambiente e pela acentuada patogenicidade em cordeiros o objetivo foi, determinar a ocorrência e caracterizar molecularmente a infecção por Cryptosporidium spp. em cordeiros na região Centro Sul do Estado de São Paulo, Brasil. Num total de 193 amostras de fezes foram coletadas de ovinos de diversas raças, machos e fêmeas, com idade de até um ano. Por meio da reação em cadeia da polimerase (nested PCR) para a amplificação de fragmentos de DNA a partir do gene da subunidade 18S do rRNA houve positividade de 15% e o sequenciamento dos fragmentos amplificados foi possível em 19 amostras. A análise das sequências obtidas mostraram que as espécies identificadas nesses animais foram Cryptosporidium xiaoi em 15 amostras, sendo o primeiro estudo de caracterização molecular desta espécie de Cryptosporidium no Brasil. Cryptosporidium ubiquitum em três amostras, e Cryptosporidium meleagridis em uma amostra, sendo estas duas últimas consideradas espécies zoonóticas.(AU)