<div>Background: Cryptosporidiosis is an emerging zoonotic protozoan disease caused by Cryptosporidium spp. The infection was reported worldwide from domestic animals and humans, including wild animals. From India, no such reports were published on Cryptosporidium infection in captive wildlife. Hence, a pilot study was conducted to report the occurrence of Cryptosporidium infection in captive wildlife of India.&nbsp; 
 
Methods: Faecal samples (n=788) were collected from 127 captive wildlife species of three zoological parks viz., Sri Venkateswara Zoological Park (SVZP), Tirupati (n=242); Indira Gandhi Zoological Park (IGZP), Visakhapatnam (n=218); Nehru Zoological Park (NZP), Hyderabad (n=328) and screened for Cryptosporidium infection. Preliminary screening of faecal samples was done for the detection of Cryptosporidium oocysts by modified Ziehl-Neelsen (mZN) staining method and the test positives were confirmed by nested PCR targeting 18S rRNA gene. Nested PCR amplicons were sequenced for determining the Cryptosporidium species. The resultant data were statistically analyzed by Fisher/Chi square, Fisher Exact test using SPSS software v 17.0. 
 
Result: In mZN staining method, 7.23 percent of isolates were found to be positive for Cryptosporidium and the highest rate of infection was detected in wildlife at NZP, Hyderabad (8.23%), followed by SVZP, Tirupati (7.44%) and IGZP, Visakhapatnam (5.50%). Cryptosporidium positive faecal samples by mZN staining were further confirmed by nested PCR and positive amplicons were sequenced for determination of Cryptosporidium species. Genetic characterization revealed five species viz., Cryptosporidium parvum; C. ryanae, C. suis, C. muris and Cryptosporidium avian genotype III. The study conclude that, Cryptosporidium infection was prevalent in the captive wildlife from the zoological parks of India and species variation was marked among the wildlife. Based on the available literature, the current study is the first of its kind on the prevalence of Cryptosporidium in captive wildlife from India.</div>

<div>Isolation and Genetic Characterization of Cryptosporidium from Captive Wildlife of India</div>

<div style="text-align: justify;">A sum total of 402 faecal samples from buffalo calves belonging to the age group of three months were collected randomly without any clinical symptom from various livestock farms and small holdings in four districts of Haryana. The presence or absence of Cryptosporidium spp. oocysts in faecal samples was confirmed by the standard microscopic examination of the modified Ziehl-Neelsen (MZN) stained faecal smears. The overall prevalence in buffalo calves was found to be 8.7%. The seasonal prevalence study revealed a maximum occurrence of Cryptosporidium infection during rainy followed by autumn, winters and minimum in summer season. Age related prevalence showed highest infection in calves between 16-30 days age which gradually decreased with increase in age. The DNA of representative positive faecal samples was extracted using standard phenol-chloroform-isoamylalcohol (PCI) method and then was subjected to nested PCR. All the extracted DNA were amplified for a 1325 bp primary fragment followed by a 834 bp nested fragment in a nested PCR of 18S rRNA gene, thereby confirming the presence of Cryptosporidium spp. The restricted enzyme digestion pattern (restricted fragment length polymorphism) of the 834 bp nested fragment using the restriction enzymes SspI and VspI confirmed the presence of C. parvum. Sequencing of Cryptosporidium DNA samples were carried out which also proved all samples to be of C. parvum. This is the first report of molecular identification of C. parvum in buffalo calves of Haryana and these Haryana isolates of Cryptosporidium parvum isolated from diarrhoeic buffalo calves are divergently related to rest of the part of the country isolates and indicates the zoonotic potential.</div>

Prevalence and molecular characterization of cryptosporidium species in buffalo calves of western Haryana

<div style="text-align: justify;">Bovine cryptosporidiosisis primarily associated with neonatal diarrhoea with higher morbidity than mortality in young calves till they attain immunological maturity. The present investigation relates to a report on the shedding of Cryptosporidium oocyst in two buffalo calves of buffalo farm, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar from 1st days up to 3 month of age at 15 days interval using simple conventional microscopy. By using formol-ether concentration technique followed by modified Ziehl-Neelsen (ZN) acid fast staining, Cryptosporidium oocysts were concentrated and identified. The Cryptosporidium oocysts appeared as reddish pink coloured bodies against a bluish/greenish coloured background at oil immersion using ZN staining kit. The maximum oocyst shedding was observed (2.3 oocyst / field) during 16 to 30 days of age. The oocyst shedding gradually decreased with increase in age afterwards clearly indicating the disease of young buffalo calves.</div>

Cryptosporidium oocyst shedding in buffalo calves in Haryana: A case study

<div style="text-align: justify;">Prevalence and molecular characterisation of Cryptosporidium species was done in kids belonging to organised and non-organised goat farms at Jabalpur. The overall prevalence of Cryptosporidium was 14.63%. The prevalence was non-significantly higher in male kids (16.16%) as compared to that of female kids (13.21%). Age wise prevalence was higher in kids up to one month age (16.13%) than that of kids upto 3 months age (13.99%). No significant difference was found in prevalence among different breeds and in kids kept in farm or field conditions. The prevalence was non-significantly higher in non-diarrhoeic kids than diarrhoeic kids. Most of the infections were of one score (76.6%). Molecular characterisation by PCR-RFLP of 18S SSU rRNA gene revealed presence of Cryptosporidium parvum species in positive faecal samples.</div><div style="text-align: justify;">&nbsp;</div>

Prevalence and molecular characterisation of Cryptosporidium spp. in goat kids&nbsp; &nbsp;

<div>Background: Cryptosporidium parvum is major protozoan parasite of both animals and humans with zoonotic significance. There is a paucity of information on its occurrence and associated risk factors in North Eastern states, India. The present study aimed at the molecular characterization of Cryptosporidium from cattle and cattle handlers in West district of Tripura and risk associates analysis. 
 
Methods: Faecal samples were randomly collected from cattle and cattle handlers (100 each), of West district, Tripura. The samples were subjected to sheather&rsquo;s sucrose flotation, modified Zeihl Neelsen (mZN) staining and PCR assay targeting the 18S SSUr-RNA gene for detection of Cryptosporidium. The polemerised chain reaction- Restriction length polymorphism (PCR-RFLP) was done to detect Cryptosporidium spp. and C. parvum genotypes. Association of the epidemiological variables in relation to host and environment were studied for comparative analysis. 
 
Result: The incidence of Cryptosporidium was 16 per cent and 7 per cent in cattle and cattle handlers, respectively. The molecular characterisation based on PCR-RFLP analysis revealed 14 C. parvum and two C. andersoni and three C. parvum in cattle handlers. All the 14 C. parvum from cattle and one C. parvum from cattle handlers belonged to bovine genotype (genotype II) and other two C. parvum from cattle handlers belonged to the human genotype (genotype I). Cryptosporidiosis in both cattle and cattle handlers were strongly associated with younger age group and diarrhoeic faecal consistency with significantly (p&lt;0.01) higher occurrence. However, significantly (p&lt;0.05) higher predominance of monsoon season in occurrence of the disease was observed in cattle and sex predominance was not observed in both cattle and cattle handlers.</div>

Molecular Characterisation of Cryptosporidium parvum among Cattle and Cattle Handlers from Tripura (India) and the Associated Risk Factors

<div>Background: Cryptosporidiosis is the disease of zoonotic importance and is considered to be third major cause of diarrhoeal disease of calves worldwide causing significant economic losses. The present study aimed to link Cryptosporidium spp. infections in dairy calves with the management practices as a major risk factor.&nbsp; 
 
Methods: Total 401 faecal samples from dairy calves were collected from eight large (100-1000 animals) and eight medium size (30-100 animals) dairy farms located in Jabalpur (Madhya Pradesh, India). Cryptosporidium oocysts were identified in faeces by modified Ziehl-Neelsen staining (mZN) and formal ether concentration techniques.&nbsp; 
 
Result: The prevalence of Cryptosporidium spp. was 28.7% in dairy calves. A higher prevalence and risk of cryptosporidiosis was associated with large farms (24.9% vs. 3.74%, OR=1.96), intensive farming system (27.7% vs. 0.99%, OR=1.86), drinking ground water (27.7% vs. 0.99%, OR=1.86), dirty farms (16.7% vs. 11.6%, OR=1.90), presence of other animals at farm (15.9% vs. 12.7%, OR=1.36), absence of deworming (14.7% vs.13.9%, OR=5.09) and diarrhoea treatment practice (16.2% vs. 12.5%, OR=1.07). Multi-animal calving area, dirty calving pen and cemented floor were also identified as risk factors. Practices like feeding colostrum within one hour of birth, @ 10% b.wt./day, weaning the calves within 24 hrs and using detergent to wash feeding utensils decreased the risk of infection from 1.4 to 2 times. Calves kept confined with their dam had greater prevalence and risk of infection (17.2% vs. 11.5%, OR=5.20). The findings highlight that proactive managemental practices are beneficial for decreasing the environmental contamination and improving calf health.</div>

Dairy Farm Management Practices as Risk Factors Linked to Cryptosporidium spp. Infection in Dairy Calves

<div>Background: The purpose of this study is to evaluate the efficacy of the treatment for the diarrhea due to Cryptosporidium, which is prevalent in lambs, via the concentrations of Serum Amyloid A (SAA) and Haptoglobin (Hp). Although there are experimental studies in this field, there are quite a few studies on the lambs which are infected naturally. 
 
Methods: In this study, stool samples were collected from the lambs that were affected with diarrhea and Cryptosporidium was detected via the rapid test kit (Bio-X Diagnostics S.A. Rochefort/Belgium). Following this procedure, 15 lambs were included in the study by examining same stool samples microscopically and 50 mg/kg of paromomycin sulphate (Huvepharma) was administered to the lambs in which Cryptosporidium spp. was detected for five days as a treatment. 10 healthy lambs formed the control group. Blood samples were taken from all lambs to measure SAA and Hp in the pre and post treatment stages. In the serums obtained from blood samples, concentrations of SAA and Hp were measured in ELISA by using commercial kits.&nbsp;&nbsp; 
 
Result: In the lambs with Cryptosporidium, concentrations of SAA and HP were as (SAA; 20,62&plusmn;4,83 ng/mL HP; 2,63&plusmn;0,77 &micro;g/mL) in pre-treatment and as (SAA;12,93&plusmn;3,27 ng/mL, Hp; 1,59&plusmn;0,73 &micro;g/mL) in post-treatment stage while they were as (SAA;10,07&plusmn;2,49 ng/mL, Hp; 1,03&plusmn;0,59 &micro;g/mL) in the control group. In the pre and post treatment stage measurements, a statistically significant difference (p&lt;0,05) in the values of SAA and Hp was found between the control group and the lambs wit diarrhea due to Cryptosporidium. It was suggested that this might have occurred as a result of&nbsp; inflammatory response against Cryptosporidium. In the light of these findings, it is asserted that routine measurements of SAA and Hp concentrations of the lambs with diarrhea due to cryptosporidiosis could be beneficial in the following up the treatment, determining the severity of the infection, choosing the right method for treatment and monitoring the efficacy of the chosen treatment method and in detecting subclinical diseases in veterinary medicine.&nbsp;</div>

Evaluating the Efficacy of the Treatment in Lambs with Cryptosporidium spp. via Saa and Haptoglobin

Background: Cryptosporidium spp. infection is one of the most common problems that occurs in kids under first month of life.The prevalence of Cryptosporidium may vary from farm to farm and region to region, depending upon herd size and animal husbandry practices.In India, although cryptosporidiosis had been reported in goat kids but Cryptosporidium spp. infection, risk factors and haematobiochemical alterations in goats&nbsp; of Madhya Pradesh are least known. 
 
Methods: A total of 290 goats d&rdquo;12 month of age belonging to unorganized goatry farms, situated in and around Rewa district of the Madhya Pradesh were attended and rectal contents were collected in 5 ml sterile screw capped plastic cryo vials during March 2023 to April 2024. The samples were screened for the presence of Cryptosporidium spp. oocysts in the faecal smear by modified Ziehl-Neelsen (mZN) method at Department of Veterinary Pathology, College of Veterinary Science and A.H., Rewa, Madhya Pradesh. To determine the haemato-biochemical parameters, 5 ml of blood samples were collected in aseptic condition from goats included in the study. 
 
Results: Out of 290 samples, twenty six (8.96%, 26/290) cases were tested positive for the Cryptosporidium oocysts. The highest prevalence was observed in kids below 1 month of age (14.6%). The Prevalence of cryptosporidium spp. was found higher in female goats (10.20 %) than male. The prevalence was found highest in goats belonged to dam of first parity (14.28%). Haematological observations in Cryptosporidium infected goats showed significantly (p&lt;0.05) lowered haemoglobin, total erythrocyte count, mean corpuscular haemoglobin and lymphocyte percentage but significantly higher total leukocyte count, neutrophil and eosinophil percentage than the healthy goats. Biochemical studies revealed significant reduction in total serum protein, albumin and globulin values in the infected goats. 
&nbsp;

Prevalence, Risk Factors and Haemato-biochemical Alterations Associated with Naturally Occurring Cryptosporidium Spp. Infection in Goats

Background: Cryptosporidium species and Giardia duodenalis are important protozoan parasites due to their wide host range and potential as zoonotic diseases. These parasites, responsible for Cryptosporidiosis and Giardiasis, are significant health concerns and have garnered attention from both the public and veterinary fields. Pets, such as dogs and cats, are susceptible to these infections and play a crucial role in transmitting these protozoans to humans. This study was conducted to determine the prevalence, species, genotypes and zoonotic significance of Cryptosporidium and Giardia in asymptomatic adult stray cats and dogs.&nbsp; 
 
Methods: Fecal samples from 75 adult stray animal, 42 dogs and 33 cats, were collected in Diyarbak&yacute;r city, southeastern Anatolia, Turkey. Direct immunofluorescence test was used to confirm the presence of G. duoenalis cysts and cryptosporidium spp. oocysts in collected fecal samples. Microscopic analysis was used to count the number of cysts/oocysts per gram as specified by the manufacturer. Molecular confirmation was done with Polymerase chain reaction (PCR) using Giardia SSU-rDNA and Cryptosporidium spp. SSU rRNA gene. The secondary PCR products of all positive samples were sequenced in one directions on an automated sequencer. Nucleotide sequence analysis was performed by BLAST alignment using the National Center for Biotechnology Information database.&nbsp; 
 
Result: The prevalence of Cryptosporidium was 7.14% in 42 cats and 9.09% in 33 dogs. The prevalence of G. duodenalis was 23.80% in cats and 54.54% in dogs. The average number of Cryptosporidium spp. oocysts per gram of cat and dog feces was 1866.3 and 1813.3, respectively. The average number of G. duodenalis cysts per gram of cat and dog feces was 1011.6 and 4786, respectively. All Cryptosporidium isolates were determined as C. canis in dogs and C. felis in cats. C. canis and C. felis isolates identified in the study, MT329018.1 and MN696800.1, AF159113.1 and KM977642.1 showed similarity with Genbank number, respectively. Assemblages A, E, C, D were found in dogs and assemblages A and E in cats in the result of the sequence of 130 bp gene fragments obtained from G. duodenalis isolates. This study sequence analysis of G. duodenalis cat isolates identified assemblages A (50%/10) and E (50%/10). In this study, assemblage E was the most common genotype isolated in dogs (38.8%), followed by Assemblage C (27.7%), Assemblage D (16.6%) and Assemblage A (15.5%). Assemblage A, which has zoonotic importance, was detected in cats and dogs, while assemblage E in cats and dogs was reported for the first time in Turkey.&nbsp;

Molecular Characterization and Zoonotic Significance of Cryptosporidium spp. and Giardia duodenalis in Asymptomatic Adult Stray Cats and Dogs in Turkey

Background: Cryptosporidium spp. and Enterocytozoon bieneusi are important enteric diarrheal pathogens commonly found in various animals and humans and have significant implications for public health. The purpose of present study was to investigate the prevalence, species and genotypes and zoonotic evaluation of the two parasitic pathogens in dogs and cats in Wenzhou, southern of China. 
&nbsp; 
Methods: 89 fecal samples collected from 51 dogs and 38 cats were used to investigate the positive rates of Cryptosporidium spp. and Enterocytozoon bieneusi using the nested polymerase chain reaction amplification on methods, species and genotypes of them were conformed by sequencing and alignment. 
&nbsp; 
Result: The positive rates of Cryptosporidium spp. and Enterocytozoon bieneusi were 7.8%(4/51), 17.6%(9/51) in dogs and 7.9%(3/38) ,10.5%(4/38) in cats, respectively. Sequence analysis revealed that Cryptosporidium canis, genotypes PtEb IX, CD9 and NED3 of E. bieneusi were confirmed in dogs, Cryptosporidium felis, Type IV in cats, respectively. C canis, C. felis and Type IV of E. bieneusi genotype belong to the zoonotic pathogens.The zoonotic species/genotype of these two pathogens identified in pets in this study indicated the potential zoonotic risk in pets-to- humans transmission. The effective interventions should be taken to prevent the spread of these enteric zoonotic pathogens. 
&nbsp;

Occurrence and Zoonotic Evaluation of Cryptosporidium spp. and Enterocytozoon bieneusi in Dogs and Cats in Wenzhou, Southern China