Evaluation of feeding different forms of therapeutic diet on the feed intake, digestibility, feed efficiency, and growth of calves experimentally infected with foot-mouth disease virus.

Oral ulcers induce acute weight loss due to anorexia in foot-and-mouth disease virus (FMDV) infected cattle. We hypothesized that providing a palatable form of a therapeutic diet (TD) in different physical forms would increase the feed intake, digestibility and restoration of body weight. A TD was formulated with 19% CP and 2.9 Mcal ME/kg on dry matter basis. Bull calves of 10-12 months with mean body weight of 123 ± 1.3 kg were experimentally infected with FMDV (n = 18) and offered one of the following three forms of the TD (n = 6/group) for 6 weeks post-FMDV infection (WPI): (i) TD in mash form (TDM) (ii) TD in cooked form (TDC) and (iii) TDC + customised nutrient supplement (TDCNS) such as Zn, Cu, Cr, Mn, and Se. The CNS was fed before the TDC. A group of uninfected control (n = 4) was fed TDM. Green fodder was offered in the afternoon. Dry matter intake (DMI) of TD and green fodder were recorded at 24 h interval till WPI 6. Body weight (BW) was recorded at weekly interval. Digestibility trial was conducted at WPI 6. The palatability of the TD was scored from 1- 4 and healing of tongue ulcers was analyzed by Kaplan-Meier survival curve. The results indicated that the physical form of TD increased the total DMI by WPI 3, which was supported by the restoration of BW and higher palatability score. The digestibility of all the proximate principles except EE was significantly higher (P < 0.05) in the groups that were fed TDC. It was concluded that feeding TD irrespective of the physical form, restored the ADG and DMI in the calves by WPI 3. Further, feeding cooked form of TD increased the digestibility in the FMDV infected calves and supplementation of CNS hastened the healing of glossal ulcers.

Heat ameliorative measures in Murrah buffalo (Bubalus bubalis) heifers during summer: effect on microclimate, thermal comfort, and behavior.

Water buffalo (Bubalus bubalis) is the mainstay of milk production in Asian countries including India. However, the hot climate of the country remains the biggest bottleneck to exploit the potential of this species. Therefore, a study was conducted to assess the effect of heat ameliorative measures on microclimate, thermal comfort, and behavior of Murrah buffalo heifers during summer. Buffalo heifers (n = 24) between the age of 15 and 20 months with a mean body weight of 363.75 ± 11.27 kg were randomly grouped into four treatments based on their age and body weight. The heifers kept in the shed without any cooling served as CON (control), while the animals of group CJ were tied with a cooling jacket. The buffalo heifers of the CJF group were subjected to the cooling jacket with forced air ventilation, whereas the intermittent (10 min at 2-h intervals) sprinkling followed by forced air ventilation was practiced in group SF between 900 and 1800 h throughout the experiment. The microclimatic variables were low in the sheds of groups CJ, CJF, and SF than the CON. The physiological responses such as rectal temperature, skin temperature, respiration rate, and pulse rate were reduced in groups CJ, CJF, and SF than the CON at 1400 h. The serum cortisol was less in the CJF and SF than those of CON group. The animals of the CJ, CJF, and SF groups spent more time for feeding and rumination with less frequent longer bouts. The extended periods of lying followed by shorter standing and idling were observed in groups CJ, CJF, and SF when compared with the CON. It may be therefore concluded that cooling jacket and intermittent sprinkling in combination with forced air ventilation might be helpful in improving the micro-environment, thermal comfort, and behavior of buffalo heifers particularly under tropical climates during summer.

Effect of physical form of the therapeutic diet on the behaviour of crossbred calves experimentally infected with foot-and-mouth disease virus.

The oral lesions associated with foot-and-mouth disease (FMD) negatively affect animal behavior, which can adversely impact animal production and welfare. Physical form of a therapeutic diet (TD) can improve the feed intake and wellbeing during the acute phase of FMD. Accordingly, we tested the effect of two physical forms of a previously developed TD on the behavior of calves experimentally infected with FMD virus (FMDV). Crossbred Holstein Friesian male calves of 10-12 months (n = 12) were experimentally infected with a virulent strain of FMDV and were offered a TD enriched with 19% CP and 2.9 Mcal ME/kg for 11 days post-FMDV infection. One group received the TD in mash form (TDM) while the other (n = 6/group) received it in cooked form (TDC). A group of four calves served as uninfected control and were fed TDM. The time spent by the calves on certain behaviours was recorded in a pre-set form from 06:00-18:00 h for 10 days from day 2-11 post-FMDV infection. The data was divided into two sessions. Session 1 (06:00-13:00 h) represented after the offering of TD, while session 2 (13:01-18:00 h) represented the data after offering green fodder. Based on exploratory data analysis, data recorded from day 2-7 post-FMDV infection was included in the final analysis. Linear mixed model was used by fitting treatment, day and their interaction as fixed effects while calf as random effect. Orthogonal contrast was applied by comparing the infected TDM with other two groups. The results revealed that the cooked form of TD improved the ingestion time, resting time, sleeping time and licking time from day 2-7 post-FMDV infection as compared with the infected TDM group. Ingestive behaviour was better in the infected TDC than that of TDM group (p < 0.01). The sleeping time was significantly high in the infected groups as compared to the uninfected TDM group (p < 0.01) till day 6 post-FMDV infection. Daily activities such as licking, standing and resting differed significantly between the infected TDM and TDC groups in session 1, but not in session 2. Urination and defecation did not differ significantly between the infected TDM and TDC groups. It was concluded that cooked form of TD remediated the effects of infection with FMDV as evidenced by improvement in the behaviour of the calves.

Microclimate modification in Murrah buffalo (Bubalus bubalis) heifers during summer: Effect on intake, growth and hematobiochemistry.

The present investigation evaluated the effect of microclimate modification on feed intake, growth performance, and hemato-biochemical profile of Murrah buffalo (Bubalus bubalis) heifers during summer. Twenty-four buffalo heifers, between 15 and 20 months of age with an average body weight of 363.75 ± 11.27 kg, were randomly assigned to 4 groups based on their age and body weight. The heifers of the T0 (control) group were kept under the shed without any cooling treatment, while the animals in group T1 were tied with a cooling jacket. Buffalo heifers of group T2 were subjected to a cooling jacket with forced ventilation and animals in group T3 were treated with intermittent sprinkling (10 min., at 2 h intervals) and forced ventilation between 9.00 AM and 6.00 PM during the experiment. The ambient temperature inside the shed and core body temperature were reduced in groups T1, T2, and T3 compared to T0. Heifers had higher dry matter intake when subjected to cooling treatments T1, T2, and T3, whereas water intake was reduced in T2 and T3 groups. The animals in T2 and T3 groups attained higher average daily weight gain, while the feed conversion was better in the T3 group compared to T0. The hematological measures such as hemoglobin, total erythrocyte count, and total leucocyte count were found higher in T0. The serum glucose, sodium, and potassium levels increasedand alkaline phosphatase activity decreased in groups T1, T2 and T3 when compared with T0. It can be concluded that the provision of intermittent sprinkling and cooling jacket in combination with forced ventilation could improve the microclimate, which in turn could enhance the performance of Murrah heifers during hot summer days in the tropics.

Characterization of arrhythmias, evaluation of cardiac biomarkers and their association with survival in calves suffering from foot-and-mouth disease.

INTRODUCTION: Foot-and-mouth disease (FMD) causes mortality in calves due to myocarditis; however, the effects of FMD virus on cardiac arrhythmogenesis and Purkinje cells are unknown. Identifying diagnostic and prognostic markers in FMD-affected calves may be useful in disease management in the endemic countries. MATERIALS AND METHODS: A total of 81 FMD-affected calves were prospectively monitored till death or recovery. Foot-and-mouth disease was diagnosed by serology and reverse transcriptase-polymerase chain reaction (RT-PCR). Electrocardiography was recorded and serum cardiac biomarkers were measured. Histopathological examination of the ventricular myocardium was carried out in the calves that died of FMD (n = 33). Apparently healthy calves (n = 15) served as control. RESULTS: Serology and RT-PCR consistently revealed that the FMD was caused by serotype O virus. Arrhythmias occurred in 62 of 81 (76.5%) FMD-affected calves, of which, ventricular premature complexes (VPCs) were the most common type (22%). The combined mortality rate due to ventricular tachycardia, polymorphic VPCs, and atrial fibrillation was 27.6%. Receiver operating characteristic curve analysis revealed that cardiac troponin I (cTnI) concentrations of ≥1.3 ng/mL were diagnostic of myocarditis with a sensitivity and specificity of 90% and 100%, respectively. Similarly, serum cTnI concentrations of <6.4 ng/mL were a good predictor of survival [odds ratio of 263; 95% confidence interval: 29-2371]. Histopathology of the myocardium revealed hyaline degeneration, necrosis, edema, mononuclear cell infiltration, and disruption by fibroblasts. Atrophy of the Purkinje cells was also present. CONCLUSIONS: FMD induces cardiac arrhythmias and Purkinje cell pathology in the calf. Portable ECG coupled with assay of serum cTnI would help in predicting survival in FMD-affected calves.

Short-term effect of foot-and-mouth disease (FMD) vaccination on the milk yield in the Deoni and crossbred cows.

Biannual vaccination of the cattle with inactivated foot-and-mouth disease (FMD) vaccine is the control strategy in endemic countries. Reduction in the milk yield is one of the main reasons for poor compliance of the cattle owners to FMD vaccination. As it can adversely affect the herd immunity, the present study aimed to quantify the losses in the milk yield post-FMD vaccination. Retrospective data on the milk yield (kg) recordings, days in milk, parity, and age at vaccination of the Deoni and crossbred cows were collected from 10 days before (-10) to 10 days after (+10) FMD vaccination (dpv). Days in milk were categorized into three stages of lactation for Deoni and crossbred cows. Age (month) was categorized into four classes. Least squares means of the milk yield were generated after adjusting for year, age, parity, and stage of lactation. Based on exploratory data analysis, the corrected milk yield records from -2 to +2 dpv for 5 years comprising 614 data points on Deoni cows (n=54) and 488 data points on crossbred cows (n=55) were used for the final analysis. Because of the correlated errors on the corrected milk yield, linear mixed model ANOVA was done by fitting dpv as fixed effect and cow as random effect, and the results revealed the effect of dpv was non-significant (P>0.05) in either breed. With respect to dpv 0, a marginal reduction of 90 g in the corrected milk yield in the Deoni cow was recorded on dpv 1, while the reduction was about 360 g on dpv 0 as compared dpv -1 in the crossbred cow. It was concluded that FMD vaccination caused a transient non-significant reduction in the milk yield in the Deoni and crossbred cows.

Factors influencing water intake in dairy cows: a review.

Water is a very critical nutrient in the diet of lactating animals as it performs several functions, viz., transfer of nutrients and excretions of waste products produced during metabolism, digestion, and body temperature regulation of all living organisms. Besides growth, other parameters like milk production, reproduction, adaptive potential, and feed consumption are also influenced by water status in feed and body reserves. Water intake in dairy cows is influenced directly or indirectly by several factors; direct factors include animal-related factors such as breed, body size, physiological stage, age, health, stress, and ecological adaptations and environmental factors, viz., season of the year, environmental temperature, humidity, wind speed, and precipitation. These factors significantly influence the drinking water intake of dairy cows. Nonetheless, different managemental practices such as rearing type, feeding practices, housing, water availability, and its quality can also affect water intake indirectly. However, overall understanding of these factors affecting water intake in dairy cows could help to improve the dairy husbandry practices and subsequently health, production, and welfare of dairy cows.

Hematological and serum biochemical profile in cattle experimentally infected with foot-and-mouth disease virus.

BACKGROUND AND AIM: Foot-and-mouth disease (FMD) is an acute viral infection affecting cloven-hoofed animals causing vesicular erosions in the oral cavity and interdigital space. The present study was undertaken to ascertain the time-dependent changes in clinical, hematological, and biochemical profiles in different breeds of cattle following experimental infection. MATERIALS AND METHODS: The animals were inoculated with 1.0×104 50% bovine tongue infectious dose (BTID50) by intradermolingual route. Clinical signs were observed, and blood/serum samples were collected at different time intervals. RESULTS: The white blood cell count declined sharply on days 7-13 and recovered on day 14 post-FMD infection. Biochemical analysis of serum markers for vital organ profile revealed no marked damage. However, a significant increase in blood urea nitrogen (BUN) value indicated pre-renal azotemia. Transient hyperthyroidism was indicated by the rise in T3 and T4 that can be correlated with a decrease in triglyceride and total cholesterol levels. In the cardiac damage assessment study, a distinct breed difference was observed wherein Malnad Gidda calves showed no cardiac damage. CONCLUSION: Except thyroid profile, BUN, and creatine kinase-myocardial band, all other serum biochemical parameters showed no significant abnormalities, whereas lymphopenia is the only hematological change and it is suggested that effective ameliorative measures should be targeted mainly on the feed/water intake, thyroid gland, and the level of lymphocytes.

Successful treatment of cardiac dysrhythmia associated with foot and mouth disease in a calf.

BACKGROUND: Foot and mouth disease (FMD), which causes myocarditis, results in 50% sudden death in the suckling calves. Occurrence of arrhythmias associated with FMD induced myocarditis in calves is not reported hitherto. The present work documents the arrhythmias associated with FMD in calf and their treatment using appropriate antiarrhythmic drugs. CASE DESCRIPTION: A three -month-old male Holstein Friesian crossbred calf naturally suffering from FMD was selected for the present study. FINDINGS/TREATMENT AND OUTCOME: Cardiac auscultation revealed grade 4 systolic murmurs and electrocardiography (ECG) showed sustained polymorphic ventricular premature complexes (PVPCs) with tachycardia on bipolar base apex lead. Apart from standard treatment, lidocaine 2% was administered at dose of 0.6 mg/kg intravenously over 15 min once a day and sinus rhythm was restored by 76 h post-treatment. Review of ECG and haematobiochemical examination revealed normal findings on 7th day of treatment. CONCLUSION: The study demonstrates the presence of sustained PVPCs with tachycardia due to FMD induced myocarditis and the successful use of lidocaine in restoring the sinus rhythm and recovery of the calf.

Behavioral pattern of Rohilkhandi kids under different feeding systems.

AIM: The present study designed to evaluate the effect of different feeding systems on the behavior of local Rohilkhandi kids. MATERIALS AND METHODS: A total of 21 growing goats (local goat of Rohilkhand region), weighing around 7-11 kg and aging 4-5 months, were used. These animals were kept in three groups. Group I was fed un-chopped green fodder in circular feeder (newly designed). Group II was fed un-chopped green fodder in linear feeder that was similar to the existing farm practice. Group III was fed chopped green fodder in linear feeder (modified version). Amount of concentrate and dry fodder fed was kept constant for all the three groups subject to equal increment in accordance with their increasing age. Adlibitum green fodder was made available to the animals. The experiment was conducted for 3 months. On-going behavior was recorded each day 4 h (2 h in the morning from 9:00 am to 11:00 am, after offering the feed, and same was repeated for 2 h in the afternoon, i.e., from 2:00 pm to 4:00 pm) was made between 9 am and 5 pm. The individual behaviors, viz., feeding, drinking, lying down, ruminating, idling, butting, pressing, pushing, frontal clashing, and physical displacement at feed barrier (active and passive: Without physical contact) of the goat were recorded using time-sampling method. Further, incidental activities such as defecation and urination were also recorded. RESULTS: Among all the groups, butting, head to head, and pushing were the common agonistic behavior found but values did not differ significantly. The pushing while feeding was relatively less in Group II (0.22±0.04 min) which differed significantly (p<0.05) from the other two groups. The idling time was found significantly (p<0.05) lower in Group II (1.68±0.21) as compared to Group I (4.67±0.52) and Group III (4.27±0.56). Time spent in rumination near the feeding trough as well as away from the feeding trough was also significantly higher in Group I (p<0.05) than the other two groups. Other minor activities, viz., defecation and urination were negligible. No stereotypic activities were observed. CONCLUSION: It was concluded that provision of un-chopped fodder in circular feeder could only simulate natural feeding behavior of goat but did not give any added advantage. Further, feeding chopped fodder in linear feeding trough lead to increased consumption and more time is spent on feeding than on agonistic behavior as compared to the other two groups.

Pathogenic and Genotypic Characterization of a Japanese Encephalitis Virus Isolate Associated with Reproductive Failure in an Indian Pig Herd.

BACKGROUND: India is endemic to Japanese encephalitis virus (JEV) and recurrent outbreaks occur mainly in rice growing areas. Pigs are considered to be the amplifying host for JEV and infection in gestating pigs results in reproductive failure. Most studies conducted on JEV infection in Indian pigs have been serological surveys and very little is known about JEV genotypes circulating in pigs. So the potential risk posed by pigs in JEV transmission and the genetic relationship between viruses circulating in pigs, mosquitoes and humans is poorly understood. METHODOLOGY/PRINCIPAL FINDINGS: This study was conducted in pigs with a history of reproductive failure characterized by stillborn piglets with neuropathological lesions. Japanese encephalitis (JE) suspected brain specimens inoculated intracerebrally into mice and Vero cells resulted in successful isolation of JEV/SW/IVRI/395A/2014. Clinicopathological observations in infected mice, demonstration of JEV antigen in brain, and analysis of the envelope protein identified the swine isolate as being neurovirulent. Phylogenetic analysis based on prM and E gene sequences showed that it belonged to genotype III. This swine isolate was closely related to JEV associated with the 2005 outbreak in India and JaoArS982 from Japan. Phylogenetic analysis of JEV strains collected between 1956 and 2014 in India categorized the GIII viruses into different clades blurring their spatial distribution, which has been discernible in the previous century. CONCLUSIONS/SIGNIFICANCE: Isolation of JEV from stillborn piglets and its close genetic relationship with viruses detected at least three decades ago in humans and mosquitoes in Japan suggests that the virus may have been circulating among Indian pigs for several decades. The close similarity between the present swine isolate and those detected in humans affected in the 2005 outbreak in Uttar Pradesh, India, suggests the need for more intensive surveillance of pigs and implementation of suitable strategies to control JE in India.

Isolation and molecular characterization of nephropathic infectious bronchitis virus isolates of Gujarat state, India.

Infectious bronchitis (IB) is a common, highly contagious, acute, and economically important viral disease of chickens caused by Infectious bronchitis virus (IBV, sp. Avian coronavirus). Five pooled tissue suspensions of 50 layer birds and one reference Massachusetts vaccine strain were inoculated into specific pathogen free (SPF) chicken egg for isolation of IBV. Reverse-transcription polymerase chain reaction (RT-PCR) was carried out using post inoculated allontoic fluid to amplify the spike (S) glycoprotein of S1 subunit of IBV. All the eggs inoculated with five pooled tissue samples and vaccine sample showed dwarfing and curling of SPF embryos indicative of IBV. All the five samples and the vaccine sample produced the expected amplicons of 466 bp by RT-PCR. The sequencing of five isolates revealed that all the five sequences were 99.09-100 % similar among themselves and showed 99.10-100 % nucleotide identity with the vaccine strain. On multiple sequence alignment it was found that our isolates were more similar at S1 subunit nucleotide level with the reference Ma5 and H120 vaccine strains than the reference Mass41 strain. The sequences of Anand isolates revealed further genetic changes in the circulating IBV in comparison to previous isolate of Gujarat as well as higher differences with the strains isolated in other states showing substantial changes at genetic level in Indian IBV isolates, which may partially explain the increasing incidences of IB in the country in spite of the vaccination.

Genotyping of major histocompatibility complex Class II DRB gene in Rohilkhandi goats by polymerase chain reaction-restriction fragment length polymorphism and DNA sequencing.

AIM: To study the major histocompatibility complex (MHC) Class II DRB1 gene polymorphism in Rohilkhandi goat using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and nucleotide sequencing techniques. MATERIALS AND METHODS: DNA was isolated from 127 Rohilkhandi goats maintained at sheep and goat farm, Indian Veterinary Research Institute, Izatnagar, Bareilly. A 284 bp fragment of exon 2 of DRB1 gene was amplified and digested using BsaI and TaqI restriction enzymes. Population genetic parameters were calculated using Popgene v 1.32 and SAS 9.0. The genotypes were then sequenced using Sanger dideoxy chain termination method and were compared with related breeds/species using MEGA 6.0 and Megalign (DNASTAR) software. RESULTS: TaqI locus showed three and BsaI locus showed two genotypes. Both the loci were found to be in Hardy-Weinberg equilibrium (HWE), however, population genetic parameters suggest that heterozygosity is still maintained in the population at both loci. Percent diversity and divergence matrix, as well as phylogenetic analysis revealed that the MHC Class II DRB1 gene of Rohilkhandi goats was found to be in close cluster with Garole and Scottish blackface sheep breeds as compared to other goat breeds included in the sequence comparison. CONCLUSION: The PCR-RFLP patterns showed population to be in HWE and absence of one genotype at one locus (BsaI), both the loci showed excess of one or the other homozygote genotype, however, effective number of alleles showed that allelic diversity is present in the population. Sequence comparison of DRB1 gene of Rohilkhandi goat with other sheep and goat breed assigned Rohilkhandi goat in divergence with Jamanupari and Angora goats.

Effect of drinking water temperature on physiological variables of crossbred dairy cattle at high altitude temperate region of Himalayas.

AIM: The objective of study was to investigate the effects of drinking water on certain physiological parameters such as heart rate (HR), respiration rate (RR), rectal temperature (RT) and, ruminal motility (RM). MATERIALS AND METHODS: The experiment was carried out on 18 farm bred lactating crossbred cows. The animals selected for the study were divided into three groups of six animals each on the basis of milk yield and parity and were allotted to three treatment group of six each such as ambient drinking water temperature at 10.25±0.28°C (ambient water, T1), drinking water temperature at 15-20°C (T2) and drinking water temperature at 35-40°C (T3). All the managemental practices were kept similar during experiment except drinking water temperatures physiological variables such as HR, RR, RT, and RM of the individual cow was measured and recorded twice in a day at 800 h and again at 1400 h two consecutive days in a week 15 min after providing drinking water. RESULT: HR and RR at morning and at evening recorded were within the normal physiological level for all the treatment groups. However, RT at morning was comparable in all the treatments whereas at evening it was significantly (p<0.01) higher for cows consuming in T2 and in T3 than cows consumed (T1). The RM during morning among the treatments were non-significant as compared to the rumen motility at evening was significantly higher for (T1) and (T2) than for cows in (T3). CONCLUSION: It can be concluded that offering warm drinking water at 35-40°C to crossbred lactating dairy cow is beneficial during winter at high altitude temperate region.

Factors affecting survivability of local Rohilkhand goats under organized farm.

AIM: To study the pattern of mortality as affected by age, season and various diseases in local goats of Rohilkhand region maintained at the Indian Veterinary Research Institute, Bareilly. MATERIALS AND METHODS: Post-mortem records of 12 years (2000-01 to 2011-12) were used, and total 243 mortality data were collected and analyzed. The causes of mortality were classified into seven major classes viz. digestive disorders, respiratory disorders, cardiovascular disorders, musculoskeletal disorder, parasitic disorders, mixed disorders (combination of digestive, respiratory, parasitic, and cardiovascular disorders) and miscellaneous disorders (cold, hypoglycemia, emaciation, endometritis, traumatic injury, etc.). RESULTS: The average mortality was 10.93%. The overall mortality was more during rainy season followed by winter and summer season. The mortality in 4-6 months of age was high (2.52%) followed by 0-1 month (2.34%) and 2-3 months (1.35%). The average mortality among adult age groups (>12 months) was 3.42%. The mortality showed declining trend with the advancement of age up to 3 months and then again increased in 4-6 months age group. The digestive diseases (3.51%) followed by respiratory diseases (1.89%) and parasitic diseases (1.48%) contributed major share to the total mortality occurred and the remaining disorders were of lesser significance in causing death in goats. There is significant (p<0.01; χ(2)=55.62) association between year with season and age with the season (p<0.05, χ(2)=16.083) found in the present study. CONCLUSION: This study confirms that overall mortality rate averaged 10.93% (ranged between 1.10% and 25.56%) over 12 years under semi-intensive farm condition. It was generally higher in rainy season. The mortality remains higher in kids particularly under 1 month of age. The digestive diseases contributed major share to overall mortality.

Estimation of Duloxetine Hydrochloride in Pharmaceutical Formulations by RP-HPLC Method.

Simple, specific, accurate and precise method, namely, reverse phase high performance liquid chromatography was developed for estimation of duloxetine HCl in pharmaceutical formulations. For the high performance liquid chromatography method, Phenomenox C-18, 5 µm column consisting of 250×4.6 mm i.d. in isocratic mode, with mobile phase containing 0.01M 5.5 pH phosphate buffer: acetonitrile (60:40 v/v) and final pH adjust to 5.5±0.02 with phosphoric acid was used. The flow rate was 1.2 ml/min and effluent was monitored at 231 nm. The retention time was 5.61 min. The method was validated in terms of linearity, accuracy and precision. The linearity curve was found to be linear over 0.25-4 µg/ml. The limit of detection and limit of quantification were found to be 0.10 and 0.25 µg/ml respectively. The proposed method was successfully used to determine the drug content of marketed formulations.