The oocyst wall is severed by means of mechanical injury or chemical agents. This study reports the percentage of in vitro sporocyst release following mechanical shaking in the presence of varying sizes of glass beads. Glass beads measured 0.5, 1, and 3 mm in diameter and were shaken with the oocysts for different times ranging from 5 sec to 5 min. Approximately 80% of sporocysts were released with 5 min of shaking in the presence of 3 mm glass beads, as well as 30 sec with 0.5 mm beads and 1 mm glass beads. The release of sporocysts of E. tenella was most efficient using 1 mm glass beads and treatment times of 30 sec to 1 min. Therefore, the use of 1 mm glass beads with 30 sec to 1 min of agitation is recommended in order to maximize sporocyst release and recovery and to improve the yield of viable sporozoites for use in biochemical, tissue culture, and immunological applications of coccidia.
Eimeria tenella/*physiology ; *Glass ; *Mechanical Phenomena ; Microspheres ; Oocysts/*physiology ; Parasitology/*methods ; *Stress, Physiological ; Time Factors

Cryptosporidium, a protozoan parasite, can cause cryptosporidiosis which is a gastrointestinal disease that can infect humans and livestock. Cattle are the most common livestock that can be infected with this protozoan. This study was carried out to determine the prevalence of Cryptosporidium infection in cattle in Kuantan, Pahang, Malaysia and to find out the association between the occurrence of infection and 3 different ages of cattle (calves less than 1 year, yearling, and adult cattle). The samples were processed by using formol-ether concentration technique and stained by modified Ziehl Neelsen. The results showed that 15.9% (24/151) of cattle were positive for Cryptosporidium oocysts. The occurrence of Cryptosporidium in calves less than 1 year was the highest with the percentage of 20.0% (11/55) followed by yearling and adult cattle, with the percentage occurrence of 15.6 % (7/45) and 11.8% (6/51), respectively. There was no significant association between the occurrence and age of cattle and presence of diarrhea. Good management practices and proper hygiene management must be taken in order to reduce the infection. It is highly important to control the infection since infected cattle may serve as potential reservoirs of the infection to other animals and humans, especially animal handlers.
Adult ; Animals ; Cattle* ; Cryptosporidiosis ; Cryptosporidium* ; Diarrhea ; Gastrointestinal Diseases ; Humans ; Hygiene ; Livestock ; Malaysia* ; Oocysts ; Parasites ; Prevalence

We investigate the resistance of Cryptosporidium (C.) parvum oocysts to commercial bleach treatment. The viability and infectivity of C. parvum oocysts suspended in 100, 50, 25, 12.5, 6.3 or 3.2% aqueous commercial bleach for 10, 30, 60, 120 or 180 min at room temperature were assessed by nucleic acid Syto-9 staining, histologic examination of ileum and infectivity to immunosuppressed neonatal C57BL/6N mice. Although the viability was decreased compared with normal oocysts, all oocysts in contact with serially diluted commercial bleach for 180 min were alive by nucleic acid dye Syto-9 staining. And, microscopic examination of ileum sections revealed developmental stages of C. parvum in all mice. The oocyst shedding patterns between mice infected with oocysts contacted with commercial bleach and normal control mice were not significantly different each other. Although commercial bleach is widely used as a bacterial and viral disinfectant, the present findings indicate that it is not an effective disinfectant for C. parvum oocysts under practical conditions. Authors conclude that, therefore, it is undesirable to recommend commercial bleach as a disinfectant for C. parvum oocysts.
Animals ; Cryptosporidium ; Cryptosporidium parvum ; Ileum ; Mice ; Oocysts ; Organic Chemicals ; Sodium Hypochlorite

Anticoccidial effects of the Plantago asiatica extract (PAE) were evaluated in chickens following oral infection with Eimeria (E.) tenella. This study was conducted on the 3-day-old chickens (n=30). Those animals were divided with 3 groups; PAE 0.1% treated/infected (n=10), PAE untreated/infected (n=10) and non-infected control (n=10). Chickens were fed a standard diet supplemented with or without PAE for 1 week prior to infection with E. tenella (10,000 sporulated oocysts per chicken). The effects of PAE on E. tenella infection were assessed by two parameters; fecal oocysts shedding and body weights gain. The PAE-fed chickens produced significantly reduced fecal oocysts (P<0.05) when compared to the E. tenella-infected group fed standard diet. Also, PAE-based diet, improved body weight loss caused by E. tenella infection. Our data demonstrated that PAE had remarkable anticoccidial activities against E. tenella. This finding might have implications for the development of anticoccidial drug. This study is the first to demonstrate anticoccidial effect of PAE on Eimeria parasites.
Animals ; Body Weight ; Chickens ; Diet ; Eimeria tenella* ; Eimeria* ; Oocysts ; Parasites ; Plantago*

Anticoccidial effects of Galla rhois (GR) extract were evaluated in chickens after oral infection with Eimeria tenella. This study was performed using 3-day-old chickens (n=30). The animals were divided into 3 groups as follows: GR 0.5%/infected (n=10), untreated/infected (n=10), and non-infected control (n=10). The chickens were fed a standard diet supplemented with or without GR for 1 week before infection with E. tenella (10,000 sporulated oocysts per chicken). The effects of GR on E. tenella infection were assessed by 2 parameters, number of fecal oocysts and body weight gain, and the results of the polymerase chain reaction (PCR). The GR-fed chickens produced significantly lower number of fecal oocysts (P<0.05) than the E. tenella-infected chickens who were fed the standard diet. In addition, GR-based diet improved the loss of body weight caused by E. tenella infection. Positive findings of PCR were identified by distinct bands in the samples of E. tenella-inoculated chickens. However, PCR analysis revealed no E. tenella oocysts in the feces of GR-fed chickens. Our data showed that GR extracts had remarkable anticoccidial activities against E. tenella. This finding might have implications for the development of novel anticoccidial drugs.
Animals ; Body Weight ; Chickens ; Coccidiosis ; Diet ; Eimeria ; Eimeria tenella ; Feces ; Oocysts ; Polymerase Chain Reaction

Anticoccidial effects of the root bark of Dictamnus dasycarpus Turcz (Rutaceae) extract (DDE) were evaluated in chickens following oral infection with Eimeria (E.) tenella. Three-day-old chickens (n=30) were assigned to three groups (control, untreated, and DDE 0.1% treated). Chickens were fed a standard diet supplemented with or without DDE for 1 week prior to infection with E. tenella (10,000 sporulated oocysts per chicken). The effects of DDE on E. tenella infection were assessed by two parameters; fecal oocysts shedding and body weights gain. The DDE-fed chickens produced significantly reduced fecal oocysts (P<0.05) when compared to the E. tenella-infected group fed standard diet. Also, DDE-based diet, improved body weight loss caused by E. tenella infection. Our data demonstrated that DDE had remarkable anticoccidial activities against E. tenella. This finding might have implications for the development of anticoccidial drug. This study is the first to demonstrate anticoccidial effect of DDE on Eimeria parasites.
Body Weight ; Chickens ; Dichlorodiphenyl Dichloroethylene ; Dictamnus* ; Diet ; Eimeria ; Eimeria tenella* ; Oocysts ; Parasites ; Rutaceae

Eimeria tenella and Eimeria maxima are important pathogens causing intracellular protozoa infections in laboratory avian animals and are known to affect experimental results obtained from contaminated animals. This study aimed to find a fast, sensitive, and efficient protocol for the molecular identification of E. tenella and E. maxima in experimental samples using chickens as laboratory avian animals. DNA was extracted from fecal samples collected from chickens and polymerase chain reaction (PCR) analysis was employed to detect E. tenella and E. maxima from the extracted DNA. The target nucleic acid fragments were specifically amplified by PCR. Feces secreting E. tenella and E. maxima were detected by a positive PCR reaction. In this study, we were able to successfully detect E. tenella and E. maxima using the molecular diagnostic method of PCR. As such, we recommended PCR for monitoring E. tenella and E. maxima in laboratory avian facilities.
Animals ; Chickens ; DNA ; Eimeria ; Eimeria tenella ; Feces ; Oocysts ; Pathology, Molecular ; Polymerase Chain Reaction

The present study was designed to assess the dynamic patterns of pro-inflammatory cytokines, including IFN-γ, TNF-α, IL-4, IL-6, acute phase protein (α1-acid-glycoprotein, AGP), and an inflammation associated factor (adenosine deaminase; ADA) following experimental caprine coccidiosis. Ten kids aging from 2 to 4 months were infected orally with 5×104 sporulated oocysts and 10 animals served as controls. Blood samples were collected in both groups before infection and at days 3, 7, 14, 21, 28, and 35 post-infection (PI), and the levels of above-mentioned factors were measured. IFN-γ, TNF-α, IL-4, IL-6, AGP, and ADA activities were significantly higher in infected animals from day 7 PI (P<0.05). In conclusion, the circulatory levels of most systemic inflammatory markers, including pro-inflammatory cytokines (IFN-γ, TNF-α, IL-4, IL-6), AGP, and ADA increased significantly starting from day 3 to day 7 PI in caprine coccidiosis.
Acute-Phase Proteins ; Aging ; Animals ; Coccidiosis* ; Cytokines ; Immunity, Innate* ; Inflammation ; Interleukin-4 ; Interleukin-6 ; Oocysts

OBJECTIVECryptosporidium spp. are prevalent globally and sheep are an important zoonotic reservoir. Little data regarding the rates of Cryptosporidium infections in ovines in China are available. This study assessed the prevalence of Cryptosporidium spp. in pre-weaned ovines from Aba Tibetan and Qiang Autonomous Prefecture in the Sichuan province of China.

METHODSA total of 213 fecal samples were collected from pre-weaned ovines and were examined microscopically (following modified acid fast staining). In addition, 18S rRNA genetic sequences were amplified from fecal samples by nested PCR and phylogenetically analyzed.

RESULTSThe prevalence of Cryptosporidium in the collected samples was at 14.6% (31/213) and four isolates identified by PCR belonged to the Cryptosporidium cervine genotype (Cryptosporidium ubiquitum) demonstrating that this species was the primary sheep species found in sheep in China.

CONCLUSIONThe present study suggested that the high incidence of Cryptosporidium in sheep poses a significant public health threat and that surveillance practices must be established to prevent zoonotic disease of humans.

Animals ; China ; Cryptosporidium ; genetics ; isolation & purification ; Feces ; parasitology ; Oocysts ; microbiology ; Polymerase Chain Reaction ; Sheep ; Weaning

OBJECTIVETo simultaneously detect viable Cryptosporidium parvum oocysts and Giardia duodenalis cysts for the purpose of reducing time and cost spent.

METHODSA duplex reverse transcription polymerase chain reaction (RT-PCR) method was newly developed.

RESULTSUsing duplex RT-PCR method for the hsp70 gene, viable (oo)cyst concentrations of 10(1) and 10(3) (oo)cysts/100 microL could be detected for C. parvum and G duodenalis, respectively. However, after heat-shock stimulation the expression of hsp70 mRNAs was detectable at 10(0) and 10(1) (oo)cysts/100 microL concentrations of C. parvum and G duodenalis, respectively. Thus, the detection sensitivity was significantly increased when the viable (oo)cysts were exposed to heat shock.

CONCLUSIONThis study describes a new duplex RT-PCR method for hsp70 gene to detect the viable (oo)cysts of the C. parvum and G duodenalis with less time consumed and at a lower cost. This newly developed duplex RT-PCR method may be used to detect these parasites not only in aquatic environments but also in clinical samples.

Cryptosporidium parvum ; isolation & purification ; Giardia ; isolation & purification ; Oocysts ; Reverse Transcriptase Polymerase Chain Reaction