• 한국임상수의학회지
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• 제21권3호
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• pp.248-252
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• 2004

DNase activity from infective larvae of the parasitic nematode Haemonchus contortus was characterized and compared to that from whole worm. DNase activity from infective larvae was detected throughout pHs 4-10, but high activity was detected under acidic conditions. The activity was not inhibited by 10 mM EDTA at pH 5.0, but was significantly inhibited at pH 7.0. The activity produced DNA, fragments with mixtures of 3'-hydroxyls (OH) and 3'-phosphates (P) at each pH with predominance of 3'-P. A unique DNase activity at 37 kDa was identified from infective larvae on zymograms. The 37 kDa DNase was detected only at pH 5.0, but not at pH 7.0, and this activity was not inhibited by EDTA at pH 5.0. These characteristics of the 37 kDa infective larval DNase resemble those of classic acidic DNases (e.g., DNase II). In contrast, 34, 36 and 38.5 kDa DNase activities were shown to be specific for whole worm. This result demonstrated that DNases in H contortus are regulated during development.

• Asian-Australasian Journal of Animal Sciences
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• 제20권1호
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• pp.7-11
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• 2007

The study was conducted on 162 adult male Haemonchus contortus of sheep collected from Avikanagar, Jaipur and Bikaner regions to diagnose the benzimidazole (BZ) resistance in H. contortus. The BZ resistance is primarily linked with the mutation in ${\beta}$-tubulin isotype 1 gene which substitute phenylalanine (Phe) into tyrosine (Tyr) at the 200 codon of the gene. An allele specific polymerase chain reaction (AS-PCR) technique was used for diagnosis of BZ resistance in H. contortus. In AS-PCR, one reverse primer (TGG 312) was used in two separate reactions with each of 2 forward primers (resistant TGG 331 and susceptible CAW 106 primer) that differed only at 3' nucleotide position. Therefore, the amplified products from resistant and susceptible parasites were produced 267 and 266 bp, respectively. A total of 162 parasites were genotyped, of which 130 parasites found homozygous resistant 'rr', 22 heterozygous 'rS' and 10 homozygous susceptible 'SS' type. The prevalence of 'rr' individuals was higher in Jaipur (98%) followed by Avikanagar (93%) and Bikaner (50%) regions. Overall, the prevalence of BZ resistant allele (r) was higher (87%) as compared to 13% of BZ susceptible allele (S).

• 대한수의학회지
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• 제44권3호
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• pp.455-462
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• 2004

Change in ${\beta}$-tubulin nucleic acid and protein sequences was the only known difference between Haemonchus contortus fenbendazole (FBZ)-resistant and -susceptible isolates. This change was sufficient to determine the pathologic effect induced by FBZ treatment. This research was initiated to investigate further differences from these two isolates. Since ${\beta}$-tubulin is involved in formation of microtubule, which has functions in secretory vesicle transport, DNase activities from excretory/secretory products (ESP) of the two isolates were compared, based on pH, sensitivity to DNase inhibitors, molecular masses and production of 3'-OH. The most significant difference detected was that a 38.5 kDa DNase activity was identified from ESP of H. contortus FBZ-susceptible isolates but not from those of H. contortus FBZ-resistant isolates. However, it was shown that the 38.5 kDa DNase is expressed with similar level of activity in intestine and whole worm of H. contortus FBZ-resistant and -susceptible isolates. This result demonstrated that the secretory transport pathway of the 38.5 kDa DNase was inhibited by unknown mechanisms, which may be related with ${\beta}$-tubulin sequence change in FBZ-resistant isolates. Other DNases of 34, 36 and 37 kDa were detected from ESP of both H. contortus FBZ-resistant and -susceptible isolates. Overall DNase activities found from ESP of these two isolates were not inhibited by 10 mM EDTA at pH 5.0, but largely inhibited by pH 7.0. In addition, DNase activities in two isolates produced DNA fragments with mixtures of 3'- hydroxyls (OH) and 3'-phosphates (P) at each pH although the 3'-end labeling ratios at pH 5.0 and 7.0 were shown different. Identification of inhibition of the 38.5 kDa DNase secretion in FBZ-resistant isolates suggests existence of further differences, in addition to ${\beta}$-tubulin sequence change, in two isolates. This shows complex effect of FBZ on H. contortus biological mechanisms.

• 대한수의학회지
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• 제44권3호
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• pp.441-448
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• 2004

DNase activity in Haemonchus contortus reproductive tissue was characterized and compared to that in whole worm. DNase activity in reproductive tissue was detected throughout pHs 4-10 with high activity under acidic conditions. The activity was not inhibited by 10 mM EDTA at pH 5.0, but largely inhibited by pH 7.0. The activity produced DNA fragments with mixtures of 3'-hydroxyls (OH) and 3'- phosphates (P) at each pH. Three distinct DNase activities were identified and had $M_rs$ of 34, 36 and 38.5 kDa in zymograms, which were distinguished according to pH requirement and sensitivity to EDTA. Among them, the 36 kDa reproductive tissue DNase had predominant activity at pH 5.0, but very weak at pH 7.0, and this activity was not inhibited by EDTA at pH 5.0. These characteristics of the 36 kDa reproductive tissue DNase resemble those of classic acidic DNases. In contrast, 36 kDa whole worm DNase activity had high activity at both pH 5.0 and 7.0. While the 36 kDa DNase activity at pH 5.0 was similar in both reproductive tissue and whole worm samples, the activity at pH 7.0 was predominantly detected in whole worm sample. This suggests that the 36 kDa whole worm DNase at pH 5.0 differs from that at pH 7.0. Thus, results indicate that the EDTA-insensitive 36 kDa DNase at pH 5.0 is specific for H. contortus reproductive tissue.

• Asian-Australasian Journal of Animal Sciences
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• 제9권4호
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• pp.421-426
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• 1996

Comparison of body weight changes of does infected with Haemonchus contortus (treatment groups 2 &3) and uninfected does (control group) was made using weekly body weight measurements over 35 weeks. The animals in treatment group 2 weighted significantly (p < 0.05) less than animals in the control group from the 5th week to the end of the experiment. Does in treatment group 3 had significantly (p < 0.05) lighter body weight than control does from the 13th week to the end of the study. The animals in control group gained an average of 0.75 kg. On the other hand, animals in infected groups 2 and 3 lost an average of 3.90 kg and 4.13 kg body weight, respectively. The animals in groups 2 and 3 also had significantly (p < 0.05) lower preslaughter and hot carcass weights than the controls.

• Asian-Australasian Journal of Animal Sciences
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• 제19권11호
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• pp.1524-1528
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• 2006

A total of 437 animals, comprising lambs aged between 3 and 12 months and adults of either sex of Muzaffarnagari sheep maintained at the Central Institute for Research on Goats, Makhdoom, Farah, Mathura, India were screened to assess the prevalence of Haemonchus contortus infection following natural infection and to identify the various factors affecting faecal egg count (FEC) and packed cell volume (PCV) of ewes and their genetic control. The relationships between FEC, PCV and body weight were also estimated. The prevalence rate for H. contortus infection in the flock under study was 15.7% indicating much lower occurrence of worm infection in lambs up to one year of age. On the other hand, a large proportion i.e., 67.7% of sheep was refractive to natural H. contortus infection. The random effect of sire significantly contributed (p<0.01) variation in log-transformed FEC (LFEC) of ewes. The season of birth had a significant (p<0.01) effect on LFEC of ewes. The lactating ewes had significantly (p<0.01) higher faecal egg counts compared to dry and pregnant ewes. The linear regression effects of the age of ewes on LFEC of animals were significant (p<0.01) in the present study. The heritabilities of LFEC, PCV and body weights of ewes during the course of infection were moderate to high in magnitude and ranged from 0.24 to 0.47. The LFEC of ewes was significantly (p<0.05) and negatively correlated with PCV at both genetic and phenotypic level. The genetic and phenotypic relationships between LFEC and body weights of ewes were -0.26 and -0.06 for this breed. The genetic correlation of PCV and body weight of ewes was positive and high (0.58) and statistically significant (p<0.05) but it was negatively correlated (-0.01) with body weight at the phenotypic level.

• Asian-Australasian Journal of Animal Sciences
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• 제10권6호
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• pp.679-682
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• 1997

A uniform group of 12 upgraded growing goats aged between 6.0 and 7.5 months were used in this study. They were divided into three groups of $T_1$, $T_2$ and $T_3$. Four animals were randomly allocated to each group. They were infected orally with three levels (0 larva, 5,000 larvae and 10,000 larvae) of infective Haemonchus contortus larvae. Before infection, all animals were housed in individual pens with concrete floors. They were provided with a uniform management. Total red blood cells (RBC) and total white blood cells (WBC) were measured by hemacytometric method. Results showed significant interaction effect of H. contortus infection and duration of infection on red blood cell counts. The RBC counts of animals in treatment groups 2 and 3 showed significantly lower values over the control group from the second fortnight to the end of the study. The overall mean RBC values of groups 1, 2 and 3 were 11.73, 9.70 and $9.12million/mm^3$ blood, respectively. H. contortus infection did not significantly influence the total leukocyte counts. Worm infection and duration of infection interaction was also absent on WBC counts. However, the time or duration of infection significantly influenced the WBC counts. Fecal egg counts showed patent infections in the infected animals which also indicated by postmortem worm counts.

• Parasites, Hosts and Diseases
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• 제59권2호
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• pp.167-171
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• 2021

Haemonchosis remains a significant problem in small ruminants. In this study, the assay of recombinase polymerase amplification (RPA) combined with the lateral flow strip (LFS-RPA) was established for the rapid detection of Haemonchus contortus in goat feces. The assay used primers and a probe targeting a specific sequence in the ITS-2 gene. We compared the performance of the LFS-RPA assay to a PCR assay. The LFS-RPA had a detection limit of 10 fg DNA, which was 10 times less compared to the lowest detection limit obtained by PCR. Out of 24 goat fecal samples, LFS-RPA assay detected H. contortus DNA with 95.8% sensitivity, compared to PCR, 79.1% sensitivity. LFS-RPA assay did not detect DNA from other related helminth species and demonstrated an adequate tolerance to inhibitors present in the goat feces. Taken together, our results suggest that LFS-RPA assay had a high diagnostic accuracy for the rapid detection of H. contortus and merits further evaluation.

• Asian-Australasian Journal of Animal Sciences
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• 제26권10호
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• pp.1446-1458
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• 2013

The study assessed the effect of dietary supplementation of leaf meal mixture (LMM) containing condensed tannins (CT) on feed intake, nutrient utilization and performance of sheep infected with Haemonchus contortus. Eighteen adult sheep of similar age and body weight ($25.03{\pm}1.52$) were included in this study and out of these, 12 sheep were infected with single dose of infective third stage larvae of H. contortus at 2,000 larvae per sheep. The experimental sheep were allocated in three different groups' i.e. negative control (NC; no infection), control (C; H. contortus infected) and treatment (T; H. contortus infected+CT at 1.5% of the DM through LMM) and the experiment was conducted for a period of 90 d. The intake of dry matter (DM), organic matter (OM) and digestibility of DM, OM, neutral detergent fibre (NDF) and acid detergent fibre (ADF) were comparable among three animal groups. However, digestibility of crude protein (CP) and ether extract (EE) were significantly (p<0.05) higher in NC group as compared to both C and T groups. Nitrogen (N) retention (g/d or % of N intake) was significantly (p = 0.038) lower in C group as compared to T and NC groups. Daily intake (g/kg $W^{0.75}$) of digestible crude protein (DCP), digestible organic matter (DOM) and total digestible nutrient (TDN) did not differ significantly (p<0.05) in the three groups. Haemoglobin (Hb) and packed cell volume (PCV) were significantly (p<0.001) higher in treatment group as compared to control. The level of Hb and PCV reduced (p<0.001) after 30 days of experimental feeding. CT significantly (p<0.001) reduced serum urea in T group as compared to NC and C groups. Serum proteins differed significantly (p<0.01) among the three groups. The activity of serum enzymes AST, ALT, ALP and LDH were also statistically non significant (p<0.05) among treatments. The weight of abomasal lymph nodes (ALN) in T group was higher (p<0.05) than in C group. Treatment group had lower (p<0.05) total worms and fecal egg count compared to control group. It may be concluded that dietary supplementation of CT through LMM significantly improved the N retention, and inhibited the different developmental stages of Haemonchus contortus in experimental sheep.

• Asian-Australasian Journal of Animal Sciences
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• 제10권4호
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• pp.435-438
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• 1997

Twelve newborn goat kids born of philippine does (3.5 and 5 years old) were used in this study. Four mother (candidate) doe goats were randomly allocated to each group. The mother does were infected orally with three levels (0, 15,000 and 30,000 larvae) of infective Haemonchus contortus larvae. Before infection all animals were housed in individual pens with concrete floors. They were provided with a uniform management. Estrus of does were synchronized using PGF2 alpha. All the animals were bred naturally by the same buck. Baby goat born of infected mother goats were divided into three groups. $T_1$, $T_2$ and $T_3$), respectively, from mother treatment groups. Birth weight and growth of goat kids born from H. contortus infected mother goats were determined. Birth weights of kids of $T_1$, $T_2$ and $T_3$ were 1.9, 1.5 and 1.2 kg, respectively. No significant (p > 0.05) differences in birth weight of kids for the 3 treatment groups were found. However, significant (p < 0.05) effects of stomach worm infection and duration of infection on liveweight gain of kids were observed. After second and third week of birth, respectively, the kids of groups 3 and 2 registered lover liveweight gains than the kids in control group. However, no significant (p > 0.05) difference in liveweight gain was found between the kids of infected mother does. Fecal egg counts of the infected mother does showed patent infections which also indicated by postmortem worm counts. However, no worm egg was found in the feces of the test kids.