The Age-Related Cryptosporidium Species Distribution in Asymptomatic Cattle from North-Western Spain

Wang Y., Cao J., Chang Y., Yu F., Zhang S., Wang R., Zhang L.

Cryptosporidium spp. and Giardia duodenalis are common gastrointestinal parasites with a broad range of hosts, including humans, livestock, and wildlife. To examine the infection status and assess the zoonotic potential of Cryptosporidium spp. and G. duodenalis in dairy cattle in Gansu, China, a total of 1414 fecal samples were collected from the rectum, with one sample collected from each individual animal. All the samples were tested using nested PCR based on the small subunit ribosomal RNA (SSU rRNA) gene of Cryptosporidium spp. and G. duodenalis. The overall infection rates of Cryptosporidium spp. and Giardia duodenalis were 4.2% (n = 59) and 1.0% (n = 14), respectively. Four Cryptosporidium species were identified: C. andersoni (n = 42), C. parvum (n = 12), C. bovis (n = 5), and C. ryanae (n = 1). In further analyses of subtypes of C. parvum isolates based on the 60 kDa glycoprotein (gp60) gene, five were successfully subtyped as IIdA19G1 (n = 4) and IIdA15G1 (n = 1). All 14 G. duodenalis isolates were identified as assemblage E using the triosephosphate isomerase (tpi) gene. The relatively low positive rates of Cryptosporidium spp. and G. duodenalis detected here and the predominance of non-human pathogenic species/assemblages of these parasites indicated their unique transmission dynamics in this area and the low level of threat posed to public health. However, continuous monitoring and further studies of these parasites should be conducted for the prevention and control of these pathogens.

Zahedi A., Ryan U.

Cryptosporidiosis, caused by parasite species of the genus Cryptosporidium, is a major diarrhoeal disease in both people and animals globally, with C. hominis and C. parvum the main species infecting humans. Environmentally robust oocysts which are shed in high numbers in the faeces of infected individuals are resistant to disinfectants, including levels of chlorine normally used in drinking water. As a result, Cryptosporidium is a major cause of waterborne and foodborne outbreaks. Interestingly, C. hominis is responsible for the majority of waterborne outbreaks typed to date with C. parvum responsible for the majority of foodborne outbreaks. No vaccine and few treatments are currently available, which has greatly limited control of this disease to date. Livestock are both an important reservoir and source of human infections and improved husbandry and management practices as well as a One Health integrated molecular typing approach across both veterinary and public health systems are essential to improve our ability to control this disease. • Update on waterborne and foodborne outbreaks caused by Cryptosporidium over the last 10 years. • Impact of cryptosporidiosis on livestock. • Evidence for livestock as a source of human infections. • Importance of a One Health integrated molecular typing approach to limit transmission.

Shaw H.J., Innes E.A., Morrison L.J., Katzer F., Wells B.

• Cryptosporidium parvum is a major cause of enteric disease in neonatal calves. • Clinically severe cryptosporidiosis in neonatal calves significantly reduces weight gain. • The impact of reduced weight gain in calves is seen over a 6 month period. Cryptosporidiosis can have a devastating effect in neonatal calves, resulting in diarrhoea, dehydration and, in severe cases, death of the animal. The disease is caused by Cryptosporidium spp. and is one of the most common causes of calf enteritis in the UK. The parasite is very difficult to remove from the farm, as the oocysts have a tough outer wall which enables the parasite to survive for several months in moist temperate environmental conditions and it is difficult to kill oocysts with common disinfectants used on a farm. If appropriate management practises are applied, the disease is usually self-limiting and most calves will recover. It has been shown, in studies with children and in lambs, that severe clinical cryptosporidiosis can result in long-term growth and cognitive impairment compared with individuals with no obvious signs of the disease. This study measured the long-term growth rate of beef calves on farm by comparing groups of animals that had suffered differing degrees of clinical severity of cryptosporidiosis as neonates. A group of 27 beef calves were enrolled in the study and monitored from birth to 6 months of age. The calves were scored for severity of cryptosporidiosis and weighed at regular intervals. The average difference in weight gain, at 6 months, between a group of calves that had severe cryptosporidiosis as neonates and a group of calves with no clinical signs of infection was 34 kg. Those calves that had experienced severe cryptosporidiosis as neonates showed a significantly reduced live weight gain compared with those calves showing no clinical signs of infection ( P = 0.034). Therefore, the impact of severe cryptosporidiosis in neonatal calves has longer term effects on weight gain and production efficiency, resulting in the parasite having a greater impact on cattle production than previously thought.

Yildirim A., Adanir R., Inci A., Yukari B.A., Duzlu O., Onder Z., Ciloglu A., Simsek E.

The prevalence of Cryptosporidium species in calves and heifers with relation to diarrhea from several herds was investigated in this study. Fecal samples were collected from 135 and 120 pre-weaned calves and 79 and 130 heifers raised in the Central Anatolia (CAR) and Mediterranean Regions (MR) of Turkey, respectively. A total of 86 post-weaned calves in CAR were also included in the study. For diagnostic comparison, all samples were examined by microscopic examination, SSU rRNA nested PCR and TaqMan real-time PCR for the presence of oocyst and Cryptosporidium DNA. In total, 102 (34.0 %) and 93 (37.2 %) of the examined samples from CAR and MR were found positive for Cryptosporidium DNA with both nested PCR and real-time PCR analyses, respectively with an overall prevalence of 35.5 %. The diagnostic sensitivity and specificity of microscopic examination were determined as 68.7 % and 100.0 % compared to molecular tools, respectively. RFLP and sequence analyses of the SSU rRNA from the PCR products revealed that 138 (70.8 %) out of 195 positive isolates were C. parvum further confirming the species-specific real-time PCR results. Among the remaining 57 (29.2 %) positive isolates, 30 (15.4 %) and 27 (13.8 %) were characterized as C. ryanae and C. bovis, respectively. C. parvum was the dominant species in pre-weaned calves especially with diarrhea while C. bovis and C. ryanae were mostly found in post-weaned calves and heifers. The sequence analyses of the gp60 gene of C. parvum isolates revealed two subtypes (IIaA13G2R1, IIaA14G1R1) belonging to zoonotic family IIa, with IIaA13G2R1 being the most common in diarrheic calves.

Åberg M., Emanuelson U., Troell K., Björkman C.

In order to investigate the infection dynamics of Cryptosporidium bovis and C. ryanae, a two-year prospective cohort study was performed on a dairy farm known to be free of C. parvum. Sixteen calves were recruited when newborn. Faecal samples were collected weekly until calves were nine weeks old, then monthly until calving or culling. Samples (n = 455) were examined by fluorescence microscopy, and when positive the species were determined by DNA sequencing. In calves up to nine weeks, C. bovis was found in 58.5% of the samples, C. ryanae in 9.2%, and both C. bovis and C. ryanae in 3.1%. The prevalence of shedding calves peaked at 87.5% in week five, which is earlier than many international studies have shown for C. bovis. The cumulative incidence of C. bovis reached 100% when the calves were five weeks old. In four calves, the species detected changed from C. bovis to C. ryanae or the other way around, and two samples were a combination of both species. Several individuals shed oocysts sporadically up to 16 months of age. The highest oocysts per gram faeces count was seen in week three (3.6 × 106 OPG). Diarrhoea was not associated with oocyst shedding.

Li F., Zhang Z., Hu S., Zhao W., Zhao J., Kváč M., Guo Y., Li N., Feng Y., Xiao L.

Bamboo rats are widely farmed in southern China for meat, but their potential in transmitting pathogens to humans and other farm animals remains unclear. To understand the transmission of Cryptosporidium spp. in these animals, 709 fecal samples were collected in this study from Chinese bamboo rats (Rhizomys sinensis) on nine farms in Jiangxi, Guangxi and Hainan provinces, China. They were analyzed for Cryptosporidium spp. using PCR and sequence analyses of the small subunit rRNA gene. Cryptosporidium parvum, C. parvum-like and C. ubiquitum-like genotypes identified were subtyped by sequence analysis of the 60 kDa glycoprotein (gp60) gene. Altogether, Cryptosporidium spp. were detected in 209 (29.5%) samples. The detection rate in samples from animals under two months of age (70.0%,105/150) was significantly higher than in samples from animals above 2 months (18.6%, 104/559; χ2 = 150.27, df = 1, P < 0.0001). Four Cryptosporidium species/genotypes were identified: C. parvum (n = 78); C. occultus (n = 1); a new genotype that is genetically related to C. ubiquitum (n = 85); and another new genotype that is genetically related to C. parvum (n = 44). Among them, C. parvum (27,610 ± 71,911 oocysts/gram of feces) and the C. parvum-like genotype (38,679 ± 82,811 oocysts/gram of feces) had higher oocyst shedding intensity than the C. ubiquitum-like genotype (2470 ± 7017 oocysts/gram of feces) and the C. occultus (1012 oocysts/gram of feces). The C. parvum identified belonged to three subtypes in two rare subtype families, including IIpA9 (n = 43), IIpA6 (n = 6) and IIoA15G1 (n = 9), while the C. parvum-like and C. ubiquitum-like genotypes generated very divergent gp60 sequences. Results of the present study suggest that bamboo rats on the study farms were infected with diverse Cryptosporidium species and divergent C. parvum subtypes, which probably had originated from their native habitats. As similar C. parvum subtypes have been recently detected in humans and farmed macaques, attentions should be paid to the potential role of these new farm animals in the transmission of zoonotic pathogens.

Santin M.

Cryptosporidium and Giardia are ubiquitous protozoan parasites that infect a broad range of vertebrate hosts, including domestic and wild animals as well as humans. Both parasites are of medical and veterinary importance. Infections with Cryptosporidium and Giardia in ruminants are associated with diarrhea outbreaks, mainly in young animals. Ruminants are potential sources of infection for humans because some species of Cryptosporidium and assemblages of Giardia duodenalis have been isolated from both ruminants and humans. Knowledge of these parasites has greatly expanded in the last 2 decades from simple microscopic observations of organisms to the knowledge acquired from molecular tools.

Xu N., Liu H., Jiang Y., Yin J., Yuan Z., Shen Y., Cao J.

Cryptosporidium is a genus of common intestinal protozoa, members of which cause diarrhea in a wide variety of hosts. Previous studies on Cryptosporidium in China have mainly focused on diarrhea sufferers, children, and immunodeficient individuals such as HIV/AIDS patients. However, the epidemiological characteristics of Cryptosporidium in the population in rural areas remain unclear. Herein, we investigated the prevalence of, and risk factors for, Cryptosporidium in rural areas of Binyang County, Guangxi Zhuang Autonomous Region, China, and genetically characterized the Cryptosporidium isolates we obtained. From August to December 2016, two villages in Binyang County, Guangxi, were sampled using a random cluster sampling method. Fresh fecal samples were collected from all eligible residents (residence time > 6 months). Molecular characterization of Cryptosporidium was carried out based on its SSU rRNA, gp60, actin and hsp70 gene sequences. Fisher’s exact test were conducted to assess the risk factors for Cryptosporidium infection. A total of 400 fecal samples were collected from 195 males (48.8%) and 205 females (51.2%). Two samples (0.5%) were positive for Cryptosporidium and were identified as C. viatorum and C. occultus respectively. Moreover, a new C. viatorum subtype XVaA3h was identified based on the sequence of the gp 60 gene. To our knowledge, this is the first report of C. viatorum and C. occultus infections in humans in China and of C. viatorum subtype XVaA3h. The findings provide important information on the prevalence of Cryptosporidium in the Chinese population, and expand the range of Cryptosporidium species known to infect people in China.

Zhao W., Zhou H., Huang Y., Xu L., Rao L., Wang S., Wang W., Yi Y., Zhou X., Wu Y., Ma T., Wang G., Hu X., Peng R., Yin F., et. al.

Wild rats (Rattus spp.) carry many zoonotic pathogens including Cryptosporidium. Due to the close proximity of rats to humans in urban environments, the potential for disease transmission is high. Cryptosporidium is a protozoan parasite which when ingested causes serious human illness. Despite its importance, genetic characterization of Cryptosporidium in wild rats in the Hainan province of China has not been performed. In this study, we analyzed the occurrence and genetics of Cryptosporidium in wild rats from Hainan, China. From December 2017 to October 2018, 150 wild rats were captured and fresh fecal material was collected from intestinal sections. Rat species were identified by PCR-based amplification and analysis of the vertebrate cytochrome b (cytb) gene. Cryptosporidium was examined by PCR amplification of the partial small subunit of ribosomal DNA (SSU rDNA). C. viatorum were subtyped by PCR analysis of the gp60 gene. A total of four rat species were identified including Asian house rats (Rattus tanezumi) (n = 46), brown rats (Rattus norvegicus) (n = 56), Edward's long-tailed rats (Leopoldamys edwardsi) (n = 38) and muridae (Niviventer fulvescens) (n = 10), with Cryptosporidium positive rates of 73.9%, 28.6%, 55.3% and 40.0%, respectively (average infection rate: 50.0%, 75/150. Sequence analysis confirmed the presence of four Cryptosporidium species and two genotypes including C. viatorum (n = 11); C. occultus (n = 2); C. muris (n = 1); and C. erinacei (n = 1); rat genotypes III (n = 13) and IV (n = 47). Three novel subtypes of C. viatorum were identified in 6 of the 11 infected Edward's long-tailed rats: XVcA2G1a (n = 4), XVcA2G1b (n = 1) and XVdA3 (n = 1). The identification of human pathogenic C. viatorum and zoonotic C. occultus, C. muris and C. erinacei, suggested that wild rats infected with Cryptosporidium pose a threat to human health. Taken together, these findings highlight the need to control the rat population in Hainan, China. The need to improve the public awareness of the risk of disease transmission from wild rats to humans is also highlighted.

Åberg M., Emanuelson U., Troell K., Björkman C.

In order to investigate the infection dynamics of Cryptosporidium bovis and Cryptosporidium ryanae, a Swedish dairy farm known to be free of C. parvum was recruited for a one-year study. Fecal samples were collected every four weeks (a total of 13 occasions) from 20 randomly selected calves younger than 65 days. A total of 238 feces samples were collected directly from the rectum and then examined with immunofluorescence microscopy; those positive for Cryptosporidium spp. oocysts were further processed by molecular species determination. Oocysts were found in 92 samples, and in 72 of these the species was successfully determined: 87.5% C. bovis, 9.7% C. ryanae and 2.8% a mix of both species. No C. parvum was found. The prevalence of shedding calves was highest at ages 4 and 5 weeks; however, the feces with the highest oocyst per gram (OPG) were registered at 2-4 weeks of age. This is earlier than what has been found in most other studies, indicating that the infection dynamics of C. bovis/C. ryanae may be affected by the presence of C. parvum. The OPG for the positive calves ranged from 200 to 1.1 × 106 (geometric mean 6.88 × 103). The youngest calf in which C. bovis was identified was 5 days old, and the youngest calf in which C. ryanae was identified was 15 days old. Calves' housing type and seasonality were not associated with differences in the shedding of oocysts. Furthermore, there was no association between the presence of diarrhea and oocyst shedding.

Feng Y., Ryan U.M., Xiao L.

Cryptosporidium species differ in host range. Parasite-host coevolution, host adaptation, and geographic segregation have led to the formation of subtype families with unique phenotypic traits within the major human-pathogenic species C. parvum and C. hominis. Transmission intensity, genetic diversity, and occurrence of genetic recombination and selective pressure have further shaped their population genetic structures. Panmixia appears to be common within the zoonotic C. parvum, especially its hypertransmissible IIaA15G2R1 subtype. Genetic recombination in C. hominis, in contrast, is more restricted to virulent subtypes, especially IbA10G2. Nonhuman primates and equine animals are commonly infected with genetically divergent C. hominis populations. Systematic studies of these and other host-adapted Cryptosporidium spp. are likely leading to improved understanding of population structures underlying various transmission patterns and intensities of Cryptosporidium.

Manyazewal A., Francesca S., Pal M., Gezahegn M., Tesfaye M., Lucy M., Teklu W., Getachew T.

A cross-sectional study was conducted to determine the prevalence and risk factors of Cryptosporidium infection and identify species of the parasite in cattle in central Ethiopia. Faecal samples, collected from 392 dairy cattle managed under intensive and extensive production system, were analyzed by the Modified Ziehl-Neelsen (MZN) microscopy, Nested PCR, PCR-RFLP and sequence analyses of the SSU rRNA gene of Cryptosporidium. The overall prevalence, the prevalence in the extensive and intensive farms was 18.6%, 11% and 21%, respectively. The infection was detected in 37.7% of the investigated farms with prevalence range of 7.4% -100%, and all of the six surveyed districts with significant (P = 0.000) prevalence difference. Restriction digestion and sequence analysis showed Cryptosporidium parvum and C. andersoni in 27% and 73% of the infections, respectively, showing an age related distribution pattern, C. parvum exclusively occurring in calves

Liang N., Wu Y., Sun M., Chang Y., Lin X., Yu L., Hu S., Zhang X., Zheng S., Cui Z., Zhang L.

AbstractTo determine the prevalence of Cryptosporidium in dairy cattle in Guangdong Province, South China, 1440 fecal samples were collected from 10 farms and screened for Cryptosporidium with PCR. The overall prevalence of Cryptosporidium was 4.38% (63/1440), and the infection rates in preweaned calves, postweaned calves, heifers and adults were 6.4% (19/297), 6.19% (33/533), 1.48% (4/271) and 2.06% (7/339), respectively. Three Cryptosporidium species, Cryptosporidium andersoni (n = 33), Cryptosporidium bovis (n = 22) and Cryptosporidium ryanae (n = 8) were detected by DNA sequence analysis of the 63 positive samples, and C. andersoni was identified as the most common species on the dairy cattle farms. In preweaned calves, C. bovis was the most prevalent species (9/19, 47.4%). In contrast, C. andersoni was the predominant species (19/33, 57.6%) in postweaned calves and the only species found in heifers and adults. The zoonotic species Cryptosporidium parvum was not detected in this study. Twenty-four C. andersoni isolates were successfully classified into three multilocus sequence typing (MLST) subtypes. MLST subtype A4,A4,A4,A1 was the predominant subtype, and MLST subtype A2,A5,A2,A1, previously found in sheep, was detected in cattle for the first time. A linkage disequilibrium analysis showed that the C. andersoni isolates had a clonal genetic population structure. However, further molecular studies are required to better understand the epidemiology of Cryptosporidium in Guangdong.

Kváč M., Vlnatá G., Ježková J., Horčičková M., Konečný R., Hlásková L., McEvoy J., Sak B.

Cryptosporidium parvum VF383 has been reported in humans, domesticated ruminants, and wild rats worldwide and described under several names including Cryptosporidium suis-like, based on its close phylogenetic relationship to C. suis. Unlike C. suis, however, it has never been detected in pigs. In the present work, C. parvum VF383 originating from wild brown rats was not infectious for piglets or calves but was infectious for laboratory brown rats, BALB/c mice, and Mongolian gerbils. The prepatent period was 4-5 days for all rodents. The patent period was longer for rats (>30 days) than other rodents (

Díaz P., Navarro E., Prieto A., Pérez-Creo A., Viña M., Díaz-Cao J.M., López C.M., Panadero R., Fernández G., Díez-Baños P., Morrondo P.

Application of molecular approaches has led to a significant progress on the knowledge of the epidemiology of Cryptosporidium spp. Nevertheless, molecular information on the occurrence of cryptosporidiosis in domestic small ruminants, especially in goats, are limited and restricted to the study of a modest number of isolates, mainly from diarrhoeic neonates. In order to determine the Cryptosporidium species present in healthy post-weaned and adult small ruminants from north-western Spain and to analyse a possible age-related distribution of species, faecal specimens were collected in sheep and goat farms without neonatal diarrhoea outbreaks the year before the sampling. Cryptosporidium spp. DNA was detected by SSU-rRNA PCR-RFLP, using restriction enzymes SspI, VspI and MboII. C. parvum and C. ubiquitum isolates were further characterized at the GP60 locus. Our results reveal that Cryptosporidium spp. is widely distributed in small ruminant farms (47.4-50.0%), although its prevalence is low in both hosts (5.9-6.0%). No significant differences in individual prevalence were detected between age groups. C. xiaoi and the zoonotic C. parvum and C. ubiquitum were identified. In sheep, C. parvum was the predominant species and its prevalence increased with age, in contrast to C. xiaoi; C. ubiquitum was an occasional finding in adults. In goats, C. xiaoi and C. ubiquitum were the most frequent species and slightly more prevalent in adults than in post-weaned kids, in contrast to C. parvum. Subtyping analysis of C. parvum isolates revealed the presence of IIaA15G2R1 and IIaA14G2R1 in sheep, whereas IIaA13G1R1 and IIdA17G1 were restricted to goats; only the C. ubiquitum XIIa subtype 3 was found. Although the prevalences detected are low, these values are probably underestimated due to, amongst others, the cross-sectional design of the study and the intermittent oocyst-excretion of post-weaned and adult small ruminants. Thus, these animals may play an important role in the appearance of cryptosporidiosis outbreaks in humans and domestic ruminant neonates and therefore should be considered as a potential threat to animal production and human health.

Li D., Li L., Yang J., He J., Zou F., Shu F.

Cryptosporidium spp. are protozoan pathogens that are widespread within mammals. In recent years, extensive molecular epidemiology studies on Cryptosporidium in dairy cattle have been conducted in Yunnan and worldwide. However, the infection status of these pathogens in beef cattle in Yunnan remains unclear. To examined the occurrence of Cryptosporidium spp. in beef cattle in Yunnan Province, China, we collected 735 fecal samples from six breeds of beef cattle in five regions of Yunnan. Nested PCR and DNA sequencing revealed the infection, species, and genotypes of Cryptosporidium spp. in these animals. The occurrence of Cryptosporidium spp. in Simmental cattle, Brahman cattle, Aberdeen Angus cattle, Yunnan Yellow cattle, Dulong cattle, and Hereford cattle was 32.9% (137/416), 3.8% (4/106), 24.4% (20/82), 3.8% (3/79), 3.2% (1/31), and 0% (0/21), respectively, with an overall rate of 22.4% (165/735). Regarding the regions, the occurrence of Cryptosporidium spp. in Boshan City, Kunming City, Lincang City, Dehong City and Xishuangbanna City was 41.8%, 28.6%, 19.4%, 6.7%, and 3.8%, respectively. In terms of age, the infection rates of Cryptosporidium spp. in pre-weaned, post-weaned, juvenile, and adult cattle were 62.1%, 52.6%, 42.7%, and 7.7%, respectively. According to sex, male cattle were more susceptible to Cryptosporidium infection (28.0%) than females (15.7%). Four Cryptosporidium species were identified in beef cattle: C. andersoni (n = 146), C. bovis (n = 11), C. ryanae (n = 7), and C. occultus (n = 1). Multilocus sequence typing analysis at the MS1, MS2, MS3, and MS16 gene loci revealed four subtype families of C. andersoni (A4A4A4A1, A5A4A4A1, A4A4A2A1, A1A4A4A1). Additionally, sequencing analysis of the 60-kDa glycoprotein gene identified three subtype families of C. bovis (XXVIc, XXVId, XXVIe) and one subtype family of C. ryanae (XXIb). These findings document the occurrence of Cryptosporidium spp. in beef cattle in Yunnan Province for the first time, providing reference data on the distribution, infection rate, species diversity, and genetic structure of these pathogens in China. To effectively reduce the prevalence of Cryptosporidium spp. in beef cattle in Yunnan, the implementation of proper sanitation management, rigorous rodent control, and farmer education programs is crucial. These integrated measures are critical for maintaining herd health, reducing economic losses, and ensuring meat safety across the province.

AbdAllah O.R., Gabre R.M., Mohammed S.A., Korayem A.M., Hussein H.E., Ahmad A.A.

Abstract Background Synanthropic filth flies thrive in human and animal habitats, posing health risks through the transmission of infectious agents. They breed on organic waste, including animal feces, making them carriers of various pathogens. In Egypt, where livestock farming is common and poor sanitation, these flies may contribute to zoonotic disease transmission. The current study investigates parasitic infections in filth flies from three livestock farms in Assiut Governorate, Upper Egypt, highlighting their role as vectors for zoonotic infections, particularly Cryptosporidium, via morphological and molecular tools. Methods A total of 12,749 flies were collected from the study sites via sweep nets. After taxonomic identification, the flies were examined microscopically for parasites using various concentration and staining techniques. Positive samples were further confirmed for infections, particularly for Cryptosporidium parasites, via nested PCR and sequence analysis targeting the COWP and SSU rRNA genes. Results This study revealed the presence of several fly species from seven dipteran families, particularly the family Muscidae, primarily Musca domestica, which presented a high parasite infestation rate of 96.6%. This study revealed a high prevalence of various protozoans and helminths in the collected flies. Cryptosporidium was the most prevalent parasite (64.4–100%), infecting all fly species. Entamoeba and Balantidium were also significant, especially in M. domestica (22.6–90.1%, 8.9–100%), Fannia canicularis (10.5–74.4%, 44.2–88.2%), and Borborillus vitripennis (11.1–50%, 37.2–91.4%). Giardia, Trichuris, and Trichostrongylidae had low to moderate prevalence in multiple fly species. Mites are commonly detected on fly exoskeletons, with high infestation rates observed in Musca domestica (77–100%) and Physiphora alceae (66.7–100%). The present study also reported sporadic infections with Trichomonas, Toxocara vitulorum, and pseudoscorpions, along with notable midge larval infestations (52.1%), mainly at site B. Parasitic infections were highest in autumn and spring, with the lowest rates in winter. Molecular identification confirmed the presence of the zoonotic species Cryptosporidium parvum and Cladotanytarsus gedanicus. Conclusion This study revealed that zoonotic parasites exist in flies and pose potential risks when they are found near humans. Cryptosporidium parvum is the prevalent parasite causing diarrhea outbreaks in animals. This is the first genetic evidence of Cladotanytarsus gedanicus midge from Upper Egypt.

Del Río M.C., Martín S., Quílez J., Vergara-Castiblanco C., Molina J.M., Ferrer O., Conde M.M., Molina J.A., Ruiz A.

Aboelsoued D., Toaleb N.I., Abdel Megeed K.N.

Abstract Cryptosporidium sp. is an obligatory intracellular apicomplexan protozoan parasite that causes a disease called cryptosporidiosis with substantial veterinary and medical importance. Therefore, this study aimed to evaluate an early diagnosis of cryptosporidiosis using the anti-Cryptosporidium parvum oocyst immunoglobulin IgG polyclonal antibodies (anti-C. parvum IgG PAbs)-based sandwich enzyme-linked immunosorbent assay (ELISA) for the detection of Cryptosporidium oocyst antigens in fecal samples of farm animals in Egypt. Further molecular identification and sequencing were performed for the detected isolates. Eight hundred and twenty fecal samples of farm animals; 102 buffalo calves, 120 cattle calves, 100 lambs and 98 goat kids, 80 buffaloes, 60 cattle, 160 sheep and 100 goats, collected from different small-scale farms and local holders were examined for cryptosporidiosis by Modified Ziehl-Neelsen (MZN) technique. The percentage of positivity was 45.1%, 50%, 20%, 18.4%, 31.25%, 38.3%, 18.8%, and 11% in buffalo calves, cattle calves, lambs, goat kids, adult buffaloes, adult cattle, sheep, and goats, respectively. Molecular identification of Cryptosporidium samples was performed based on COWP gene, revealing the isolates: GenBank: OQ121955.1, OR029973.1 and PP316107.1 which were identical to the C. parvum and GenBank: PP316108.1 and OR029972.1 which were identical to C. hominis and C. andersoni, respectively. Then, C. parvum oocysts were used for preparation of antigens and rabbit immunization. Anti-C. parvum IgG PAbs were purified and characterized by SDS-PAGE and then labeled with horseradish peroxidase (HRP). Anti-C. parvum IgG PAbs in-house sandwich ELISA was prepared, then tested this ELISA on 820 samples and compared results with MZN microscopical examination and a commercial sandwich ELISA kit. In this study, in-house sandwich ELISA scored higher sensitivity of 98%, 100% specificity, validity 99% and relative agreement 98.6% than (92%, 90%, 91% and 91.4%) of MZN and (96%, 95%, 95.5% and 95.7%) of coproantigen commercial sandwich ELISA kit, respectively. Moreover, we used PCR to evaluate the positivity of in-house sandwich ELISA results, and the total PCR positive samples were 263 out of 268 sandwich ELISA positive samples (98.13%). In conclusion, the prepared Anti-C. parvum IgG PAbs based sandwich ELISA offered a simple and accurate diagnostic method for cryptosporidiosis in the fecal samples of different species of farm animals in Egypt with high sensitivity (98%) and specificity (100%). Further studies on this Anti-C. parvum IgG PAbs may help also in the protection against cryptosporidiosis.

López-Novo C., Couso-Pérez S., Prieto A., Díaz-Cao J.M., García-Dios D., López-Lorenzo G., Remesar S., Ares-Mazás E., López C., Morrondo P., Gómez-Couso H., Díaz P.

Rideout H., Cook A.J., Whetton A.D.

Cryptosporidium species are parasitic organisms of vertebrates with a worldwide distribution. They have an important impact globally upon human and animal health, and livestock productivity. The life cycle of these species is complex and difficult to disrupt to improve human health, animal health, food security and economic growth. This may contribute to the fact that no new treatment strategy has been widely accepted or applied in livestock for years. Here we consider the natural history of these parasites, their biochemistry and economic impact. Using recent developments in understanding these parasites we then consider viable and affordable approaches to enhancing control of their effects on livestock. These are based on advances in drug discovery, omics research and artificial intelligence applications to human and veterinary medicine that indicate putative new therapeutic approaches.

Buchanan R., Matechou E., Katzer F.D., Tsaousis A.D., Farre M.

AbstractBackgroundThe protozoan parasiteCryptosporidiumis the causative agent of a severe diarrhoeal disease, called cryptosporidiosis.Cryptosporidiumspecies are capable of infecting a wide range of hosts including humans and livestock. In cattle, cryptosporidiosis is now one of the most important causes of neonatal scour globally, either as a sole agent or co-infecting with other pathogens. Cryptosporidiosis is considered globally endemic, with a prevalence ofCryptosporidiumin stool samples from 13% to 93% in European cattle. This disease has a significant economic burden, with costs associated with veterinary diagnosis, medication, increased labour, animal rearing and supplemental nutrition as well as being associated with reduced long-term growth rate in calves, causing huge economic losses in livestock industry. Moreover, cattle act as a zoonotic reservoir forCryptosporidium parvum, a species that is capable of infecting humans as well. As such, monitoring the prevalence ofCryptosporidiumin cattle is important due to the public health risk and financial burden the clinical disease causes.MethodsPublications reporting on the prevalence ofCryptosporidiumin cattle were collected from PubMed and Google Scholar. Information regarding the species ofCryptosporidiumin positive samples, the genotype ofC. parvumfound in samples, and the diarrhoeic status of the cattle was collected where available. A total of 279 publications were collected for this meta-analysis from six continents and 65 countries to provide an estimation for global bovineCryptosporidiumprevalence.ResultsA 25.5% global prevalence ofCryptosporidiuminfection was reported, withC. parvumbeing the most frequently identified species, particularly the IIa subfamily. Diarrhoea was reported in 14,141 cattle samples, of which 36.0% tested positive forCryptosporidium. Regarding symptoms, we found that in countries reporting over 50% of diarrhoeic positive cattle,C. parvumwas the most common species.ConclusionsContinued monitoring and reporting ofCryptosporidiumin cattle are crucial for both public health and economic reasons. Consequently, efforts should focus on underreported regions and the development of control measures to reduce prevalence and limit zoonotic transmission.

Gao J., Zhou L., Zhang A., Hou M., Liu X., Zhang X., Wang J., Wang X., Bai X., Jiao C., Yang Y., Lan Z., Qiu H., Wang C.

Crytosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi are important diarrheal pathogens with a global distribution that threatens the health of humans and animals. Despite cattle being potential transmission hosts of these protozoans, the associated risks to public health have been neglected. In the present study, a total of 1155 cattle fecal samples were collected from 13 administrative regions of Heilongjiang Province. The prevalence of Cryptosporidium spp., G. duodenalis, and E. bieneusi were 5.5% (64/1155; 95% CI: 4.2–6.9), 3.8% (44/1155; 95% CI: 2.7–4.9), and 6.5% (75/1155; 95% CI: 5.1–7.9), respectively. Among these positive fecal samples, five Cryptosporidium species (C. andersoni, C. bovis, C. ryanae, C. parvum, and C. occultus), two G. duodenalis assemblages (E and A), and eight E. bieneusi genotypes (BEB4, BEB6, BEB8, J, I, CHS7, CHS8, and COS-I) were identified. Phylogenetic analysis showed that all eight genotypes of E. bieneusi identified in the present study belonged to group 2. It is worth noting that some species/genotypes of these intestinal protozoans are zoonotic, suggesting a risk of zoonotic disease transmission in endemic areas. The findings expanded our understanding of the genetic composition and zoonotic potential of Cryptosporidium spp., G. duodenalis, and E. bieneusi in cattle in Heilongjiang Province.

Golomazou E., Mamedova S., Eslahi A.V., Karanis P.

Cryptosporidiosis is a significant contributor to global foodborne and waterborne disease burden. It is a widespread cause of diarrheal diseases that affect humans and animals worldwide. Agricultural environments can become a source of contamination with Cryptosporidium species through faecal material derived from humans and animals. This review aims to report the main findings of scientific research on Cryptosporidium species related to various agricultural sectors, and highlights the risks of cryptosporidiosis in agricultural production, the contamination sources, the importance of animal production in transmission, and the role of farmed animals as hosts of the parasites. Agricultural contamination sources can cause water pollution in groundwater and different surface waters used for drinking, recreational purposes, and irrigation. The application of contaminated manure, faecal sludge management, and irrigation with inadequately treated water are the main concerns associated with foodborne and waterborne cryptosporidiosis related to agricultural activities. The review emphasizes the public health implications of agriculture concerning the transmission risk of Cryptosporidium parasites and the urgent need for a new concept in the agriculture sector. Furthermore, the findings of this review provide valuable information for developing appropriate measures and monitoring strategies to minimize the risk of infection.

G S.A., A. P.N., Kasaralikar V.R., Sandeep H., G R.B., S P.B., T R.

Abstract The cryptosporidiosis is the major protozoal disease of calves. The overall prevalence of cryptosporidiosis was found to be 14.63 per cent in 205 diarrhoeic calves. The mZN staining method was economical and sensitive tool for diagnosis of cryptosporidiosis compared to other microscopic examinations. The positive samples were subjected to nPCR analysis and confirmed the presence of Cryptosporidium spp. in the 30 suspected samples at 834bp on agarose gel electrophoresis. The nPCR products were sequenced and revealed the presence of C.parvum in the two samples. The phylogenetic tree of sequenced samples suggested that the present samples were in close relation with isolates of Punjab (India), Bangladesh and China. Haemogram indicated that the calves were marginally anaemic and dehydrated along with corresponding alterations in erythrocyte indices. There was significant increase in the total leukocyte count in the present investigation. Hypoproteinemia and hypoalbuminemia were observed with corresponding decrease in the A:G ratio although the alterations were marginal. Histologically, there was desquamation of epithelial cells on intestinal mucosa and atrophy of villus crypts thus reducing the crypt to villi ratio. Nitazoxanide and azithromycin were equally effective in eliminating the infection with proper supportive therapy, however treatment with azithromycin was cost effective.

Rzeżutka A., Kaupke A.

AbstractCattle cryptosporidiosis is noted worldwide with varied frequency of infection prevalence depending on geographical, environmental and husbandry factors. In this study, the prevalence of Cryptosporidium infections in cattle was determined on the basis of molecular results obtained by testing 1601 faecal samples collected from calves up to 4 months of age housed in all Polish provinces from 2014 to 2018. Detection and identification of Cryptosporidium species was performed at the 18 small subunit ribosomal RNA (18S rRNA) locus by conducting PCR–RFLP analysis of the amplified DNA fragments. The prevalence of Cryptosporidium infections in the cattle population was 45.3% (CI 95%: 42.8–47.7; 725/1601). The infected animals were housed on 233/267 (87.3%) of monitored farms with regional prevalence ranging from 27.8 to 62%. The restriction pattern of 18S rRNA amplicons for positive samples was characteristic of C. parvum, C. bovis, C. ryanae, C. andersoni, and unexpectedly also of C. baileyi and C. suis. Infections of C. bovis and C. ryanae prevailed in the studied cattle population relegating C. parvum to third in prevalence. Likewise, mixed infections caused by C. bovis and C. ryanae as well as C. parvum and C. bovis were observed. A relationship between the infecting parasite species and animal breed was found. For instance, C. parvum prevailed in Black and White lowland breed, C. ryanae in Limousine cattle and C. andersoni in dairy animals of mixed dairy breeds. Furthermore, differences in prevalence of particular parasite species between cattle breeds were also shown.

Gomes-Gonçalves S., Palmeira J.D., Ferreira H., Santos-Silva S., Mesquita J.R.

Enteropathogenic parasites are of significant concern for public health due to their zoonotic potential and their impact on human and animal health. In this study, we investigated their occurrence and characterized these enteropathogens in asymptomatic domestic ruminants from Portugal. A total of 302 stool samples were collected from cattle (n = 166), sheep (n = 73), and goats (n = 63) in various regions of Portugal and tested for Cryptosporidium spp., Giardia duodenalis, Enterocytozoon bieneusi, Blastocystis sp., and Balantioides coli by PCR. The occurrence of Cryptosporidium spp. was found to be 12.7% (8/63, 95% confidence interval [CI]: 5.65–23.5) in goats; however, no sample was found to be positive for Cryptosporidium spp. in cattle and sheep. For E. bieneusi, 6.35% (4/63; 95%CI: 1.76–15.47) of goats were found to be positive; however, no cattle or sheep were found to be positive. Blastocystis sp. was found in sheep (9.59%; 7/73; 95% [CI]: 0.394–18.76) and goats (12.70%; 8/63; 95% [CI]: 5.65–23.50) but none was found in cattle. No positive results for G. duodenalis or B. coli were detected in this study. This study provides essential baseline information for understanding the silent shedding and epidemiology of these enteropathogens in Portugal, contributing to overall livestock health and related occupational safety. Raising awareness among consumers, veterinarians, and farm owners is crucial to minimize the risk of transmission and promote effective disease control strategies.

Figueiredo A.M., Köster P.C., Dashti A., Torres R.T., Fonseca C., Mysterud A., Bailo B., Carvalho J., Ferreira E., Hipólito D., Fernandes J., Lino A., Palmeira J.D., Sarmento P., Neves N., et. al.

Enteric protozoan parasites Giardia duodenalis, Cryptosporidium spp., and, to a lesser extent, the ciliate Balantioides coli are responsible for severe human and animal intestinal disorders globally. However, limited information is available on the occurrence and epidemiology of these parasites in domestic, but especially wild species in Portugal. To fill this gap of knowledge, we have investigated G. duodenalis, Cryptosporidium spp., and B. coli occurrence, distribution, genetic diversity, and zoonotic potential by analyzing 756 fecal samples from several wild carnivores (n = 288), wild ungulates (n = 242), and domestic species (n = 226) collected across different areas of mainland Portugal. Overall, infection rates were 16.1% (122/756; 95% CI: 13.59–18.96) for G. duodenalis and 2.7% (20/756; CI: 1.62–4.06) for Cryptosporidium spp., while no ungulate sample analyzed yielded positive results for B. coli. Giardia duodenalis was found across a wide range of hosts and sampling areas, being most prevalent in the Iberian lynx (26.7%), the Iberian wolf (24.0%), and the domestic dog (23.9%). Cryptosporidium spp. was only identified in wild boar (8.4%), red fox (3.4%), Iberian lynx (3.3%), red deer (3.1%), and Iberian wolf (2.5%). Sequence analysis of G. duodenalis determined zoonotic assemblage A (subassemblage AI) in one roe deer sample, canine-specific assemblages C and D in Iberian wolf, red fox, and domestic dog, and ungulate-specific assemblage E in wild boar, sheep, cattle, and horse. Six Cryptosporidium species were identified: C. scrofarum in wild boar, C. canis in the Iberian wolf and red fox, C. ubiquitum in red deer and wild boar, C. felis in the Iberian lynx, and both C. ryanae and C. occultus in red deer. Giardia duodenalis and Cryptosporidium spp. coinfections were observed in 0.7% (5/756) of the samples. This is the first, most comprehensive, and largest molecular-based epidemiology study of its kind carried out in Portugal, covering a wide range of wild and domestic hosts and sampling areas. The detection of zoonotic Cryptosporidium spp. and G. duodenalis subassemblage AI demonstrates the role of wild and domestic host species in the transmission of these agents while representing a potential source of environmental contamination for other animals and humans.

Gattan H.S., Alshammari A., Marzok M., Salem M., AL-Jabr O.A., Selim A.

AbstractCryptosporidium is one of the causative parasitic agents that causes gastrointestinal diseases in calves. The parasite poses a zoonotic risk to immunocompromised individuals and children. Thus, this study aimed to determine the prevalence of Cryptosporidium infection in calves in three Egyptian governorates situated in Nile Delta and assess the associated risk factors. The Cryptosporidium oocysts were detected in 81 out of 430 calves (18.84%). In addition, the univariant analysis showed that age, feeding source, hygienic status, presence of diarrhea and contact with other animals were significantly (P < 0.05) associated with Cryptosporidium prevalence in calves. Furthermore, the risk factors related with Cryptosporidium prevalence were age (OR 1.96, 95%CI 0.97–3.94), feeding on milk and pasture (OR 2.07, 95%CI 1.15–3.72), poor hygienic condition (OR 2.25, 95%CI 1.28–3.94), presence of diarrhea (OR 2.47, 95%CI 1.23–4.96) and contact with other domestic animals (OR 2.08, 95%CI 1.24–3.50). In addition, the PCR assay targeting 18srRNA showed that the most prevalent species among calves was C. parvum. Although additional researches are required to understand the most effective steps that farmers and veterinary professionals should take to decrease the occurrence of Cryptosporidium infection.

OLIVEIRA M.C., OLIVEIRA D.R., GONÇALVES N.R., ARANTES U.M.

ABSTRACT Protozoa of the genus Cryptosporidium are parasites capable of causing animal rearing losses due to diarrhea outbreaks. This review aimed to determine the prevalence of and potential risk factors for bovine cryptosporidiosis in Brazil. The search was carried out in the databases Science Direct, Scopus, Web of Science, and SciELO. The search strategy used the descriptors “Cryptosporidium,” “cryptosporidiosis,” “cattle,” “calves,” and “Brazil.” Nine studies conducted in different Brazilian states between 2008 and 2019 were selected. The age of the evaluated animals ranged from 0 to >36 months. The main risk factors were the low age of the animals, collective pens, presence of diarrhea, contact with other animals, and dairy breed. The average prevalence rate was 30.51%. The species identified were C. ryanae, C. parvum, and C. bovis. Optical microscopy was used as a diagnostic method for oocysts in fecal samples and to determine the species, and PCR was used in three studies. The average prevalence of Cryptosporidium spp. in cattle in Brazil was 30.51%, ranging from 10.2 to 62.5%. The main risk factors found were young animals, the presence of animals with diarrhea, contact with other animals, dairy breeds, and collective pens.