PCR amplification and sequence analyses of ITS-1 rDNA from Cryptosporidium andersoni in dairy cattle

2006 ◽  
Vol 100 (5) ◽  
pp. 1135-1138 ◽  
Author(s):  
Rongqiong Zhou ◽  
Guoqing Li ◽  
Shumin Xiao ◽  
Yanxun Xia ◽  
Yuanzhong Guo
Keyword(s):  
Dairy Cattle ◽  
Pcr Amplification ◽  
Sequence Analyses ◽  
Cryptosporidium Andersoni

188 DETECTION BY POLYMERASE CHAIN REACTION OF NEOSPORA CANINUM FROM OVUM PICKUP AND UTERINE FLUSHING FLUIDS FROM DAIRY CATTLE IN MEXICO

2010 ◽  
Vol 22 (1) ◽  
pp. 252
Author(s):  
A. F. Marques ◽  
C. G. Ortiz ◽  
M. R. Lima ◽  
E. L. Zanella ◽  
L. Rangel ◽  
...  
Keyword(s):  
Dairy Cattle ◽  
Neospora Caninum ◽  
Pcr Amplification ◽  
Positive Control ◽  
Negative Control ◽  
Genomic Region ◽  
Sample Collection ◽  
Caninum Infection ◽  
Blood Lymphocytes ◽  
Negative Results

Neospora caninum, an intracellular protozoon, causes encephalomyelitis in dogs (Bjerkas I et al. 1984 Zentralblat fur Parasitenkunde 70, 271-274). For the past decade, neosporosis has been a main cause of abortion in dairy cattle worldwide (Anderson M et al. 2000 Anim. Reprod. Sci. 60-61, 417-431; Dubey JP 2003 Korean J. Parasitology 41, 1-16). Vertical transmission has been indicated as an important way of spreading neosporosis (Hall CA et al. 2005 Vet. Parasitology 31, 231-41); thus, we investigated whether the protozoon could be transferred by embryo production techniques. Blood samples were collected from 92 dairy cows with history of reproductive failure and abortion within the previous 90 days at 7 dairy farms in Tizayuca, Mexico. For serology evaluation, a commercial indirect ELISA kit (Civtest Bovis Neospora, Laboratories Hipra S.A, Girona, Spain), yielded 46.74% (43/92) positive results, 46.74% (43/92) negative results, and 6.52% (6/92) suspicious to N. caninum infection. Thirteen positive cows were chosen for uterine flush (UF), ovum pickup (OPU), and a blood sample collection. Lymphocytes from blood and cells within the UF and OPU collection fluids were collected after centrifugation and DNA was extracted. All samples were tested for the presence of N. caninum by PCR, using primers and protocols that amplified a 275-bp fragment of the genomic region (5-GGGTGAACCGAGGGAGTTG-3 and 5-CCTCCCAATGCGAACGAAA-3). The N. caninum vaccine (Bovilis® NeoGuard, Intervet, Santiago Tianguistenco, Mexico) was used as a positive control and water as a negative control. Uterine flush could not be obtained from 1 cow. From 13 cows seropositive to N. caninum, only 38% were positive to PCR from blood lymphocytes. In contrast, PCR amplification was obtained from OPU cell sediment in 92.31% (12/13) and in 33.33% (4/12) of UF. Of these 12 OPU- and 4 UF-positive samples, only 5 and 3 of their corresponding blood lymphocytes were positive. Our results using uterine and follicular fluid were contradictory to those published by Moskwa et al. (2008 Vet. Parasitology 158, 370-375) where oocytes and embryos were evaluated. These results indicate that N. caninum is present in the ovary and uterine lumen of the cows, suggesting a possible risk of neospora transmission during oocyte and embryo collection and transfer techniques. UNAM and UPF.

Cryptosporidium andersoni is the predominant species in post-weaned and adult dairy cattle in China

2011 ◽  
Vol 60 (1) ◽  
pp. 1-4 ◽  
Author(s):  
Rongjun Wang ◽  
Guangpeng Ma ◽  
Jinfeng Zhao ◽  
Qingbin Lu ◽  
Helei Wang ◽  
...  
Keyword(s):  
Dairy Cattle ◽  
Predominant Species ◽  
Cryptosporidium Andersoni

Molecular epidemiology of Cryptosporidium spp. in dairy cattle in Guangdong Province, South China

Parasitology ◽  
2018 ◽  
Vol 146 (1) ◽  
pp. 28-32 ◽  
Author(s):  
Nan Liang ◽  
Yayun Wu ◽  
Mingfei Sun ◽  
Yankai Chang ◽  
Xuhui Lin ◽  
...  
Keyword(s):  
Dairy Cattle ◽  
South China ◽  
Guangdong Province ◽  
Common Species ◽  
Infection Rates ◽  
Genetic Population ◽  
Cryptosporidium Andersoni ◽  
First Time ◽  
Cattle Farms ◽  
Prevalent Species

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.

scholarly journals Dynamics of hepatitis C virus NS5A quasispecies during interferon and ribavirin therapy in responder and non-responder patients with genotype 1b chronic hepatitis C

2005 ◽  
Vol 86 (4) ◽  
pp. 1067-1075 ◽  
Author(s):  
Francesc Puig-Basagoiti ◽  
Xavier Forns ◽  
Ivana Furčić ◽  
Sergi Ampurdanés ◽  
Mireia Giménez-Barcons ◽  
...  
Keyword(s):  
Chronic Hepatitis ◽  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Amino Acid ◽  
Genetic Distance ◽  
Antiviral Agents ◽  
Pcr Amplification ◽  
Rt Pcr ◽  
Sequence Analyses ◽  
Synonymous Substitutions

The quasispecies nature of hepatitis C virus (HCV) may have important implications concerning resistance to antiviral agents. To determine whether HCV NS5A quasispecies composition and dynamics are related to responsiveness to combined interferon (IFN) and ribavirin therapy, extensive sequence analyses of cloned RT-PCR amplification products of HCV-1b NS5A quasispecies of sequential isolates from 15 treated (nine sustained responders and six non-responders) and three untreated patients were performed. Accumulation of mutations in NS5A during therapy was relatively frequent in the V3 domain, but unusual elsewhere. Amino acid changes were the result of the imposition of minor variants that were already present before treatment and always occurred within the first week of therapy. Before treatment, the complexity and diversity of quasispecies were lower in isolates from responders than in those from non-responders, particularly in the V3 domain, where differences in nucleotide entropy (0·35 vs 0·64, P=0·003), genetic distance (0·0145 vs 0·0302, P=0·05) and non-synonymous substitutions (0·0102 vs 0·0203, P=0·036) were statistically significant. These differences became more apparent during treatment, because complexity and diversity remained stable or tended to increase in non-responders, whereas they tended to decrease in responders. These observations suggest that the composition and dynamics of HCV NS5A quasispecies, particularly in the V3 domain, may play a role in the response to combined IFN/ribavirin therapy.

scholarly journals Prevalence and Multilocus Genotyping of Giardia lamblia in Cattle in Jiangxi Province, China: Novel Assemblage E Subtypes Identified

2020 ◽  
Vol 58 (6) ◽  
pp. 681-687
Author(s):  
Sen Li ◽  
Yang Zou ◽  
Xue-Liang Zhang ◽  
Ping Wang ◽  
Xiao-Qing Chen ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Beef Cattle ◽  
Dairy Cattle ◽  
Nested Pcr ◽  
Giardia Lamblia ◽  
Pcr Amplification ◽  
Epidemiological Data ◽  
Jiangxi Province ◽  
Southeastern China ◽  
First Time

<i>Giardia lamblia</i> is a common enteric pathogen associated with diarrheal diseases. There are some reports of <i>G. lamblia</i> infection among different breeds of cattle in recent years worldwide. However, it is yet to know whether cattle in Jiangxi province, southeastern China is infected with <i>G. lamblia</i>. The objectives of the present study were to investigate the prevalence and examine the multilocus genotypes of <i>G. lamblia</i> in cattle in Jiangxi province. A total of 556 fecal samples were collected from 3 cattle breeds (dairy cattle, beef cattle, and buffalo) in Jiangxi province, and the prevalence and genotypes of <i>G. lamblia</i> were determined by the nested PCR amplification of the beta-giardin (<i>bg</i>) gene. A total of 52 samples (9.2%) were positive for <i>G. lamblia</i>. The highest prevalence of <i>G. lamblia</i> was detected in dairy cattle (20.0%), followed by that in beef cattle (6.4%), and meat buffalo (0.9%). Multilocus sequence typing of <i>G. lamblia</i> was performed based on sequences of the <i>bg</i>, triose phosphate isomerase and glutamate dehydrogenase loci, and 22, 42, and 52 samples were amplifiable, respectively, forming 15 MLGs. Moreover, one mixed <i>G. lamblia</i> infection (assemblages A and E) was found in the present study. Altogether, 6 novel assemblage E subtypes (E41*-E46*) were identified for the first time. These results not only provided baseline data for the control of <i>G. lamblia</i> infection in cattle in this southeastern province of China, but also enriched the molecular epidemiological data and genetic diversity of <i>G. lamblia</i> in cattle.

scholarly journals First mecC and mecA Positive Livestock-Associated Methicillin Resistant Staphylococcus aureus (mecC MRSA/LA-MRSA) from Dairy Cattle in Malaysia

2020 ◽  
Vol 8 (2) ◽  
pp. 147 ◽  
Author(s):  
Erkihun Aklilu ◽  
Hui Ying Chia
Keyword(s):  
Staphylococcus Aureus ◽  
Dairy Cattle ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Meca Gene ◽  
Pcr Amplification ◽  
Diffusion Method ◽  
Methicillin Resistant ◽  
Unpasteurized Milk ◽  
Antimicrobial Susceptibility Tests ◽  
Pcr Targeting

Livestock associated Methicillin resistant Staphylococcus aureus (S. aureus) (LA-MRSA) was reported to be zoonotic and may transmit to farmers and veterinarians. The objectives of this study were to investigate the occurrence of LA-MRSA from dairy cattle and to evaluate the antimicrobial resistance profiles of the isolates. A total of 63 milk and 32 nasal swab samples were randomly collected from dairy cattle. The samples were processed to isolate S. aureus, MRSA and LA-MRSA using both phenotypic and molecular methods using PCR. The confirmed S. aureus isolates were cultured on oxacillin resistant screening agar base (ORSAB) to detect MRSA and the isolates were further confirmed by PCR targeting the mecA gene. Detection of the novel mecA gene, mecC gene was conducted by PCR amplification. The antimicrobial susceptibility tests were conducted using disc diffusion method. Results revealed 17/95 (17.89%) and 15/95 (15.79%) were positive for mecA and mecC genes respectively. Out of the 15 mecC positive isolates, 12 were positive for both mecA and mecC. The MRSA isolates showed multidrug resistance. The findings showed high prevalence of mecC-positive LA-MRSA in Malaysia and highlight the public health risks to people that may come in contact with the carrier animals or those who may consume unpasteurized milk products from these animals.

Prevalence and multilocus genotyping of Cryptosporidium andersoni in dairy cattle and He cattle in Xinjiang, China

2016 ◽  
Vol 44 ◽  
pp. 313-317 ◽  
Author(s):  
Meng Qi ◽  
Rongjun Wang ◽  
Bo Jing ◽  
Fuchun Jian ◽  
Changshen Ning ◽  
...  
Keyword(s):  
Dairy Cattle ◽  
Multilocus Genotyping ◽  
Cryptosporidium Andersoni

scholarly journals DETECTION OF SPECIFIC CAUSES OF TUBERCULOSIS IN THE DAIRY CATTLE OF BANGLADESH AGRICULTURAL UNIVERSITY BY SEQUENCING AND SEQUENCE ANALYSIS

2017 ◽  
Vol 15 (1) ◽  
pp. 13-20
Author(s):  
A. Yasmin ◽  
M. Z. Hossain ◽  
U. K. Rima ◽  
T. Ruba ◽  
M. A. H. N. A. Khan
Keyword(s):  
Nucleic Acid ◽  
Sequence Analysis ◽  
Dairy Cattle ◽  
Bacterial Species ◽  
Pcr Amplification ◽  
Tuberculin Test ◽  
Acid Base ◽  
Zoonotic Risk ◽  
Nucleic Acid Base ◽  
Tuberculin Tests

Tuberculosis (TB) is a chronic disease of man and animal. In this study intradermal tuberculin test, necropsy, histopathology, polymerase chain reaction (PCR) and sequencing techniques were used to diagnose specific cause of TB in dairy cattle of Bangladesh Agricultural University, Mymensingh. Intradermal tuberculin tests were carried out on randomly selected 100 dairy cattle. Tuberculin test positive cattle (N=05) were examined at necropsy and granulomas in lungs was seen in three cattle. Caseous necrosis and swelling of lymphnodes was seen in prescapular (N=01) and mesenteric (N=02) lymphnodes. In a case nodular lesion was seen in lungs and mesenteric lymphnodes. Portion of infected lungs and lymphnodes were snap frozen, extracted genomic DNA and PCR protocols was adapted targeting MPB83 gene. Result of PCR showed amplification of 600bp fragments in five cases. The MPB83 gene although specific for M. Bovis, the gene is less abundantly expressed by M. tuberculosis. To differentiate infectivity due to M. Bovis and M. Tuberculosis, two more PCR were adapted targeting pncA and oxyR genes. Out of five cattle tested in PCR all samples generated pncA specific 185bp and oxyR gene specific 270bp amplicons. The sequencing of MPB83, pncA and oxyR genes were carried out. Results of sequencing did not show mutation in MPB83 gene. Sequencing of pncA gene showed replacement of nucleic acid base (guanine to cytosin) in position 169 in cattle no. 5. Similarly, sequence analysis of oxyR gene (n=05) showed replacement of nucleic acid base (adenine to guanine) in position 285 in cattle no. 5. The cattle no. 5 was confirmedly infected with M. Tuberculosis and rest of the cattle were infected with M. Bovis. The tuberculin tests, necropsy, histopathology and PCR amplification of MPB83 gene may not contribute species specific detection of Mycobacterial infectivity in cattle. Sequencing and sequence analysis of pncA and oxyR genes found to differentiate infectivity in cattle due to M. Bovis and M. Tuberculosis.  Both of the bacterial species are extremely zoonotic and dairy cattle of Bangladesh Agricultural University were infected with both M. Bovis and M. Tuberculosis. It needs to tests all the dairy cattle twice in a year with tuberculin tests and dispose test reactors in order to minimize zoonotic risk.

Effects of Retinoic Acid and 1,25-Dihydroxyvitamin D3 on IFN-gamma Secretion by Mononuclear Leukocytes from Nulliparous and Postparturient Dairy Cattle

2000 ◽  
Vol 70 (3) ◽  
pp. 92-101 ◽  
Author(s):  
Burim Ametaj ◽  
Brian Nonnecke ◽  
Ronald Horst ◽  
Donald Beitz
Keyword(s):  
Retinoic Acid ◽  
Dairy Cattle ◽  
Dairy Cows ◽  
Dairy Cow ◽  
Pokeweed Mitogen ◽  
Mononuclear Leukocytes ◽  
Combined Effects ◽  
Ifn Gamma ◽  
Dihydroxyvitamin D ◽  
1,25 Dihydroxyvitamin D

Individual and combined effects of several isomers of retinoic acid (RA) and 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) on interferon-gamma (IFN-gamma) secretion by blood mononuclear leukocytes (MNL) from nulliparous and postparturient Holstein cattle were evaluated in vitro. In the first experiment, effects on incubation period (24 to 72 hours) and time of supplementation (0 to 32 hours) with all-trans, 9-cis, 13-cis-, and 9,13-dicis-RAs (0 to 100 nM) on IFN-gamma secretion by pokeweed mitogen (PWM)-stimulated (0 and 10 mug/ml) MNL from nulliparous cattle were evaluated. In the second experiment, MNL from postparturient cows (bled at 0, 2, 4, and 16 days postpartum) were stimulated with PWM (0 and 10 mug/ml) in the presence of RA isomers (9-cis- or 9,13-dicis-RA; 0 to 100 nM), 1,25-(OH)2D3 (0 to 100 nM), or with combinations of these metabolites. The results show that individual isomers of RA had no effect on IFN-gamma secretion by PWM-stimulated MNL from nulliparous or postparturient cows. Furthermore 1,25-dihydroxyvitamin D3 inhibited IFN-gamma secretion by MNL from nulliparous and postparturient dairy cows; however, the degree of inhibition was greater when 9-cis- and 9,13-dicis-RA were also present in the cultures. Finally mononuclear leukocytes from postparturient dairy cows produced substantially less IFN-gamma than did MNL from nulliparous cattle. It is concluded that retinoic acids individually did not affect the capacity of leukocytes from dairy cattle to secrete IFN-gamma. This result is in marked contrast to studies in monogastric species indicating that RAs inhibit IFN-gamma secretion by peripheral blood T cells. Inhibition of IFN-gamma secretion by 1,25-(OH)2D3 was potentiated by 9-cis- and 9,13-di-cis-retinoics acids, suggesting that an excess of dietary vitamins A and D may compromise further the naturally immunosuppressed postparturient dairy cow. Additional research is necessary to determine if the combined effects of these metabolites on IFN-gamma secretion represent an increased susceptibility of the dairy cow to infectious diseases during the periparturient period. Lower secretion of IFN-gamma by MNL from postpartutient dairy cows, relative to nulliparous cattle, suggests that recently-calved cows are naturally immunosuppressed.

Sign in / Sign up

Export Citation Format

Share Document