Evaluation of fecal smear methods for research on Cryptosporidium spp. oocysts in the feces of dairy calves

The objective of this study is to compare the direct fecal smear (DFS) and centrifugal sedimentation (CS) methods in the detection of Cryptosporidium spp. oocysts in fecal samples of dairy calves. One hundred and fourteen fecal samples were collected from calves aged up to six months from 10 dairy farms located in Palotina and Francisco Alves, Paraná, Brazil. The microscopic analysis revealed the presence of Cryptosporidium spp. oocysts in 51.75% (59/114) of the samples in both methods. In CS, 48.25% (55/114) of the samples were positive, while in DFS slides, only 6.14% (7/114) were positive. Only 4 samples were positive exclusively in DFS. To ensure that there were no false-negative results in the microscopic analysis, the 55 samples that were negative in both DFS and CS were selected for molecular analysis using the nested PCR (nPCR). Of these 55 samples, 24% (13/55) were positive and forwarded for sequencing part of the genome, which made it possible to identify C. parvum, C. bovis and C. ryanae. Besides the characterization of the Cryptosporidium species, it was possible to identify bacteria of the genus Acinetobacter interfering directly in the analyzed samples. The microscopic analysis also revealed higher sensitivity when CS was used to make the fecal smears. However, some samples that were negative in this technique had positive PCR results. Thus, molecular analysis is indicated to confirm cases of Cryptosporidium spp. Further studies are necessary to prove the specificities of the used primers since the results obtained in nPCR were positive for the protozoan but, when genetic sequencing was performed, Acinetobacter spp. was identified.

Literature Review: Coinfection in Young Ruminant Livestock—Cryptosporidium spp. and Its Companions

The protozoan Cryptosporidium parvum is one of the major causative pathogens of diarrhoea in young ruminants; therefore, it causes economic losses and impairs animal welfare. Besides C. parvum, there are many other non-infectious and infectious factors, such as rotavirus, Escherichia coli, and Giardia duodenalis, which may lead to diarrhoeic disease in young livestock. Often, more than one infectious agent is detected in affected animals. Little is known about the interactions bet-ween simultaneously occurring pathogens and their potential effects on the course of disease. In this review, a brief overview about pathogens associated with diarrhoea in young ruminants is presented. Furthermore, information about coinfections involving Cryptosporidium is provided.

Prevalence of cryptosporidiosis in goats in central Algeria

This study was aimed at determining the prevalence of cryptosporidiosis in goats in central Algeria. A total of 605 samples of goat faeces were collected from several mixed farms (sheep and goats) in the regions of Médéa, Djelfa, and Ain Defla. Cryptosporidium spp. was detected using the Ziehl-Nelseen technique modified by Henriksen and Polhenz. Cryptosporidium spp. was detected in 103 samples (17.02%). Females accounted for 69.90% of positive samples while males accounted for 30.09% of positive samples (P<0.05). The most infested animals were those aged between 2 months and 7 years (69.99%). Animals under 15 days of age were also very vulnerable to infection (59.25%) (P<0.05); Young goats with diarrhoea infested with Cryptosporidium were noticed in 25% of samples. The infestation rate during autumn was 56.31%, and 43.68% in spring (P<0.05). Post-weaned goats are potential carriers and shedders of the parasite and young are vulnerable to the infection. To conclude, further studies are needed on a larger sample and in other areas to improve our epidemiology knowledge of cryptosporidiosis in goats.

Molecular identification of Cryptosporidium baileyi in Muscovy ducks (Cairina moschata domesticus) in free-range production systems

ABSTRACT: Cryptosporidiosis is considered an infection with impact on animal health. It has been associated with high morbidity and mortality rates, leading to significant economic losses to the poultry industry. This study investigated the presence of Cryptosporidium spp. in domestic ducks of family Anatidae (Cairina moschata) from two rustic commercial aviaries located in the city of Rio de Janeiro, Brazil. A total of 315 fecal samples were collected from domestic ducks in two different areas (N=186 in area A and N=129 in area B). The microscopic analysis was conducted using a sugar centrifugal flotation technique for the identification of Cryptosporidium spp. oocysts, followed by PCR/sequencing analyses of the partial sequence of the 18S rDNA gene to determine the Cryptosporidium species. Of the 315 samples collected, only 10 (186/5.38%) from area A were positive for Cryptosporidium. The nucleotide sequence and phylogenetic analyses identified that all samples were identical (100%) and belonged to Cryptosporidium baileyi species, which is closely related to gastric species and of importance in animal health.

Effets De L’extrait Aqueux Du Moringa Oleifera Sur L’abondance Des Oocystes De Protozoaires Intestinaux Des , Usées D’origine Hospitalière À Yaoundé-Cameroun

L’accès à l’eau potable demeure une très grande préoccupation en Afrique sub-saharienne, plus particulièrement dans les agglomérations où les populations sont confrontées à la pollution, l’insuffisance d’hygiène d’assainissement et au manque de méthodes appropriées de traitement à l’échelle familiale. De ce fait, l’intégration de procédés biologiques de traitement des eaux pourrait être une alternative durable dans l’amélioration de la qualité des eaux de boisson et des eaux déversées dans l’environnement après usage. La présente étude menée de juin 2019 à avril 2020 porte sur l’optimisation de l’efficacité des feuilles de Moringa oleifera dans le traitement des eaux usées d’origine hospitalière. Ces eaux ont été traitées avec différentes concentrations (100mg/l, 200mg/l, 300mg/l, 400mg/l, 500mg/L et 600mg/l) de l’extrait aqueux de Moringa oleifera. Le suivi des paramètres physicochimiques et parasitologiques à différents temps de décantation a permis de déterminer les conditions optimales et d’évaluer l’efficacité des traitements. Il en ressort qu’en fonction des caractéristiques des eaux, les temps de décantation compris entre 24-36 h sont apparus suffisants pour éliminer la presque totalité des matières en suspension et des colloïdes. Pour le même temps de décantation, l’abattement parasitologique était en moyenne de l’ordre de 99 % pour Isospora spp, 94 % pour Cryptosporidium spp, 98 % pour Cyclospora spp et 96 % pour les oocystes de Sarcocystis spp. Au cours du traitement, la baisse de certains paramètres physicochimiques tels que la réduction des teneurs en matières organique biodégradables exprimée par l’oxydabilité et la hausse d’autres paramètres comme le pH, la conductivité électrique et des Solide Totaux Dissout ont facilité la réduction des parasites durant le traitement. Ces résultats indiquent qu’en fonction des concentrations initiales, le traitement des eaux brutes avec l’extrait aqueux des feuillets de Moringa oleifera améliore considérablement la qualité des eaux. Access to safe drinking water remains a major concern in sub-Saharan Africa, especially in urban areas where people are confronted with pollution, poor hygiene, inadequate sanitation and lack of appropriate treatment methods at the household level. Therefore, the integration of biological water treatment processes could be a sustainable alternative in improving the quality of drinking water and water discharged into the environment after use. The present study conducted from June 2019 to April 2020 focuses on optimising the efficiency of Moringa oleifera leaves in the treatment of hospital wastewater. These waters were treated with different concentrations (100mg/l, 200mg/l, 300mg/l, 400mg/l, 500mg/L and 600mg/l) of the aqueous extract of Moringa oleifera. The monitoring of physico-chemical and parasitological parameters at different decantation times made it possible to determine the optimal conditions and to evaluate the effectiveness of the treatments. The results showed that, depending on the characteristics of the water, settling times between 24-36 hours appeared to be sufficient to eliminate almost all the suspended matter and colloids. For the same settling time, parasitological removal was on average around 99% for Isospora spp, 94% for Cryptosporidium spp, 98% for Cyclospora spp and 96% for Sarcocystis spp oocysts. During the treatment the decrease values of some physicochemical parameters such as the reduction of biodegradable organic matter content expressed by oxidability and the increase of other parameters such as pH, electrical conductivity and Total Dissolved Solids facilitated the reduction of parasites during the treatment. These results indicate that, depending on the initial concentrations, treatment of raw water with the aqueous extract of Moringa oleifera leaflets significantly improves water quality.

Detection of Cryptosporidiosis in Dogs of Veterinary Clinics in Surabaya City Using Acid-Fast Staining and PCR

The need for maintaining pets, such as dogs, is increasing along with the human population. When individuals keep dogs as their pets, they must be aware of disease transmission from dogs. One of the disease agents transmitted from pets to their owners is Cryptosporidium spp. causing cryptosporidiosis. The aim of the present study was to detect Cryptosporidium spp. infection in dogs through a fecal examination using the acid-fast staining method (Ziehl Neelsen) confirmed with the molecular examination of Polymerase Chain Reaction (PCR). Detection of Cryptosporidium sp. in feces of dogs was set up by using an acid-fast staining method. Positive results of the acid-fast staining were further confirmed using PCR. Polymerase Chain Reaction used primary AB210854 specific to the Cryptosporidium canis and S139-S141 genes which were specific primary for the Cryptosporidium parvum gene. Results of the acid-fast staining showed that 80% of the samples (40 samples from total samples) were infected with Cryptosporidium spp. Further detection using PCR showed that four samples were positive for Cryptosporidium canis infection, and two samples showed positive results of Cryptosporidium parvum infection. Dog samples were mostly infected with Cryptosporidium spp. including Cryptosporidium canis and Cryptosporidium parvum through a fecal examination using acid-fast staining and PCR. Keywords: Acid-fast staining, Cryptosporidium spp., Dogs, PCR

Pathogens Removal in a Sustainable and Economic High-Rate Algal Pond Wastewater Treatment System

This study evaluates the efficiency of a sustainable technology represented in an integrated pilot-scale system, which includes a facultative pond (FP), a high-rate algal pond (HRAP), and a rock filter (RF) for wastewater treatment to produce water that complies with the Egyptian standards for treated wastewater reuse. Still, limited data are available on pathogen removal through HRAP systems. Thus, in this study, the performance of the integrated system was investigated for the removal of Escherichia coli (E. coli), coliform bacteria, eukaryotic pathogens (Cryptosporidium spp., Giardia intestinalis, and helminth ova), somatic coliphages (SOMCPH), and human adenovirus (HAdV). Furthermore, physicochemical parameters were determined in order to evaluate the performance of the integrated system. The principal component analysis and non-metric multidimensional scaling analysis showed a strong significant effect of the integrated system on changing the physicochemical and microbial parameters from inlet to outlet. The mean log10 removal values for total coliform, fecal coliform, and E. coli were 5.67, 5.62, and 5.69, respectively, while 0.88 log10 and 1.65 log10 reductions were observed for HAdV and SOMCPH, respectively. The mean removal of Cryptosporidium spp. and Giardia intestinalis was 0.52 and 2.42 log10, respectively. The integrated system achieved 100% removal of helminth ova. The results demonstrated that the system was able to improve the chemical and microbial characteristics of the outlet to acceptable levels for non-food crops irrigation. Such findings together with low operation and construction costs of HRAPs should facilitate wider implementation of these nature-based systems in remote and rural communities. Overall, this study provides a novel insight into the performance of such systems to eliminate multiple microbial pathogens from wastewater.

Cross-Border Investigations on the Prevalence and Transmission Dynamics of Cryptosporidium Species in Dairy Cattle Farms in Western Mainland Europe

Cryptosporidium is an apicomplexan parasitic protist, which infects a wide range of hosts, causing cryptosporidiosis disease. In farms, the incidence of this disease is high in animals such as cows, leading to extensive economic loss in the livestock industry. Infected cows may also act as a major reservoir of Cryptosporidium spp., in particular C. parvum, the most common cause of cryptosporidiosis in these animals. This poses a risk to the trading of livestock, to other farms via breeding centres, and to human health. This study is a part of a global project aimed at strategies to tackle cryptosporidiosis. To reach this target, it was essential to determine whether prevalence was dependent on the studied countries or if the issue was borderless. Indeed, C. parvum occurrence was assessed across dairy farms in certain regions of Belgium, France, and the Netherlands. At the same time, the animal-to-animal transmission of the circulating C. parvum subtypes was studied. To accomplish this, we analysed 1084 faecal samples, corresponding to 57 dairy farms from all three countries. To this end, 18S rRNA and gp60 genes fragments were amplified, followed by DNA sequencing, which was subsequently used for detection and subtyping C. parvum. Bioinformatic and phylogenetic methods were integrated to analyse and characterise the obtained DNA sequences. Our results show 25.7%, 24.9% and 20.8% prevalence of Cryptosporidium spp. in Belgium, France, and the Netherlands respectively. Overall, 93% of the farms were Cryptosporidium positive. The gp60 subtyping demonstrated a significant number of the C. parvum positives belonged to the IIa allelic family, which has been also identified in humans. Therefore, this study highlights how prevalent C. parvum is in dairy farms and further suggests cattle as a possible carrier of zoonotic C. parvum subtypes, which could pose a threat to human health.

Effect of season and geographic location in the United States on detection of potential enteric pathogens or toxin genes in horses ≥6-mo-old

We investigated the effects of season and geographic location on detection of nucleic acids of potential enteric pathogens (PEPs) or their toxins (PEP-Ts) in feces of horses ≥6-mo-old in the United States. Results of 3,343 equine diarrhea PCR panels submitted to Idexx Laboratories for horses >6-mo-old were reviewed. Submission months were grouped into 4 seasons, and states were grouped into 4 geographic regions. Logistic regression was performed to assess effects of season and region on detection rates of PEPs and PEP-Ts. Agresti–Coull CIs were determined. Detection rate of Salmonella enterica was higher in the South in summer compared to all other regions, and was also higher in the South in fall compared to the Midwest and Northeast. The Neorickettsia risticii detection rate was lower during summer in the West and higher in fall in the Midwest. Detection of Cryptosporidium spp. was lower during spring, summer, and winter in the West. Differences were not identified for detection rates of Clostridioides difficile, Clostridium perfringens, Lawsonia intracellularis, Rhodococcus equi, equine rotavirus, and equine coronavirus. Overall, our data support seasonal and regional differences in detection rates of S. enterica, N. risticii, and Cryptosporidium spp. in horses ≥6-mo-old in the United States.