R. Somvanshi

Detection of BPV-1 and -2 and quantification of BPV-1 by real-time PCR in cutaneous warts in cattle and buffaloes.

Bovine cutaneous warts (CWs) were investigated in Northern India. Of 49 cases, 44 were recorded in cattle and 5 in buffaloes. These animals had mild to moderate grade infections. Grossly, cases of CWs appeared to be of exophytic type, however, different types of growth patterns were observed. A total of 26 biopsies (cattle 21 and buffaloes 5) from CWs-affected animals studied histopathologically were diagnosed as exophytic and cauliflower-like fibropapilloma 13, exophytic and dome-shaped fibropapilloma 5, occult and/or fibroblastic type papilloma 3, cauliflower-like papilloma 3, endophytic fibropapilloma 1 and fibroma 1. On PCR analysis, 11 CWs and 2 normal skin samples showed BPV-1, -2 mixed infections. A rapid, sensitive and reliable real-time SYBR Green PCR test to detect BPV-1, BPV-2 and to quantify BPV-1 was developed. Results of amplification and dissociation plot of real-time PCR revealed that six samples were BPV-1 positive, eight were BPV-2 positive and six were positive for both BPV-1 and -2. CWs samples from different dairy farms testing positive for BPV-1 by PCR assay were also positive using Quantitative real-time SYBR Green PCR assay. For the first time, mixed infection of BPV-1 and -2 was detected in India and BPV-1 load was quantified by real-time SYBR Green PCR assay.

Detection and quantification of bovine papilloma virus type 2 (BPV-2) by real-time PCR in urine and urinary bladder lesions in enzootic bovine haematuria (EBH)-affected cows.

This study was conducted with the objectives of detecting bovine papillomavirus type 2 (BPV-2) in urine samples and urinary bladder lesions in bovines using polymerase chain reaction (PCR) and real-time PCR-based molecular diagnostic tests, and quantifying BPV-2 in urinary bladder lesions especially in enzootic bovine haematuria (EBH)-affected animals. BPV-2 viral DNA was detected in urine samples (50%) and urinary bladder tissue (68.6%). Cloning and sequencing results showed a close homology with other Indian BPV-2 sequences. Quantitative real-time PCR (SYBR Green assay) showed that the BPV-2 load was low and similar irrespective of inflammatory or neoplastic lesions in the bladder. It was concluded that BPV-2 DNA is frequently present in urine and urinary bladder lesions in cows in an EBH endemic region and virus load was low in urinary bladder lesions.

Papillomatosis in buffaloes: a less-known disease.

Scant information is available on papillomatosis in buffaloes, and it is an almost unknown disease. It has been described from India, Italy and Turkey. Buffalo papillomatosis occurs in cutaneous and mucosal forms. Cutaneous papillomatosis is manifested as cutaneous wart (CW) and teat papilloma types. The condition is known to be caused by bovine papillomaviruses (BPV)-1 and 2 and their mixed infections. Buffalo CWs are experimentally transmissible to hamsters, cattle as well as buffaloes. Once BPV establishes infection in buffaloes, infection spreads from buffalo to buffalo, without cattle intermediary. Histologically, CWs are mostly diagnosed as fibropapillomas. The mucosal form occurs as urinary bladder tumours similar to enzootic bovine haematuria which is also associated with bracken fern infested areas. BPVs are yet to be demonstrated in teat papillomas and urinary bladder tumours of buffalo cases. Papillomatosis in buffaloes is a little-known disease, but it is a separate infectious ailment of buffaloes and deserves more attention by researchers.

Prebiotic potential of Jerusalem artichoke (Helianthus tuberosus L.) in Wistar rats: effects of levels of supplementation on hindgut fermentation, intestinal morphology, blood metabolites and immune response

BACKGROUND Many studies have been conducted using purified prebiotics such as inulin or fructooligosaccharides (FOS) as nutraceuticals, but there is very little information available on the prebiotic potential of raw products rich in inulin and FOS, such as Jerusalem artichoke (JA; Helianthus tuberosus L.). The present experiment aimed to evaluate the prebiotic effects of JA tubers in rats. RESULTS Seventy-two Wistar weanling rats divided into four groups were fed for 12 weeks on a basal diet fortified with pulverized JA tubers at 0 (control), 20, 40 and 60 g kg(-1) levels. Enhanced cell-mediated immunity in terms of skin indurations (P = 0.082) and CD4+ T-lymphocyte population (P = 0.002) was observed in the JA-supplemented groups compared with the control group. Blood haemoglobin (P = 0.017), glucose (P = 0.001), urea (P = 0.004) and calcium (P = 0.048) varied favourably upon inclusion of JA. An increasing trend (P = 0.059) in the length of large intestine was apparent in the JA-fed groups. The tissue mass of caecum (P = 0.069) and colon (P = 0.003) was increased in the JA-supplemented groups, accompanied by higher (P = 0.007) caecal crypt depth. The pH and ammonia concentrations of intestinal digesta decreased and those of lactate and total volatile fatty acids increased in the JA-fed groups. CONCLUSION The results suggest that JA had beneficial effects on immunity, blood metabolites, intestinal morphometry and hindgut fermentation of rats.

Identification of bovine papilloma virus 10 in teat warts of cattle by DNase-SISPA

Papilloma viruses are detected and identified by PCR with consensus primers designed from human papilloma virus sequences. These and other primers could not detect papilloma virus in bovine teat wart samples despite repeated attempts. DNase-SISPA, a metagenomic method for identifying viruses, could identify bovine papilloma virus type 10 in bovine teat warts. The sequence comparison between consensus primers and bovine papilloma virus type 10 sequences revealed many differences between consensus primers and BPV-10 sequences. We suggest, DNase-SISPA may be used as an alternate method for papilloma virus diagnosis, in cases where PCR fails to identify papilloma viruses.

Spontaneous Cutaneous Papillomatosis in Yaks and Detection and Quantification of Bovine Papillomavirus-1 and -2

Seven clinical cases of cutaneous papillomatosis in yaks were studied in Arunachal Pradesh, India. Sporadic, single or a chain of multiple varying size warts appeared around the eyes or on the body. Predominant site of warts was around eyes. Histopathologically, these cases were diagnosed as fibropapilloma. It was confirmed by the detection of BPV-1 and BPV-2 or their mixed infection by PCR and sequencing. Quantitative SYBR Green real-time PCR detected comparatively lower viral DNA copy number in cutaneous warts (CWs). Cases of CWs and its causative agent as bovine papillomavirus (BPVs) are reported for the first time in yaks.

Detection of Bovine Papilloma Viruses in Wart‐Like Lesions of Upper Gastrointestinal Tract of Cattle and Buffaloes

In present investigation, etiopathological characterization of upper gastrointestinal tract (GIT) tumours of cattle and buffaloes was undertaken. A total of 27 GIT wart-like lesions in rumen, reticulum, mouth and oesophagus of cattle and buffaloes revealed the presence of small nodular to larger spherical or slender growths with thin base present on mucosa and ruminal pillar. Histopathologically, these cases were diagnosed as fibropapilloma/papilloma. This is the first world record on ruminal papillomatosis in buffaloes. Ruminal warts of cattle and buffaloes revealed the presence of BPV-5, -1 & -2, which is the first report of presence of these BPVs in the ruminal warts from India. Quantitative real-time PCR revealed that DNA samples of different GIT wart-like lesions contained varying amount of BPV DNA copy numbers. Immunohistochemistry revealed that the PCNA and Ki67 immunopositivity was present in the basal and spinosum layer of the fibropapilloma/papilloma, indicating these as the cellular proliferation site. In conclusion, the present investigation revealed that BPV-5, -1 & -2 are associated with certain ruminal wart-like lesions/growths in cattle and buffaloes, and the basal and spinosum layer of the ruminal fibropapilloma/papilloma were cellular proliferation sites.

Pathological study of non-neoplastic urinary bladder lesions in cattle and buffaloes: a preliminary report.

A total of 236 urinary bladders (94 cattle and 142 buffaloes) collected from Bareilly, Uttar Pradesh, India, were studied for spontaneous lesions. These adult animals belonged to Institute’s organized dairy farm and rural areas in the Rohilkhand region of Uttar Pradesh. Grossly, congestion, hemorrhages, and cystoliths in urinary bladders were diagnosed. Histopathologically, the major conditions diagnosed were acute cystitis, 44 (18.64%), including, congestion, hemorrhages, sub-acute cystitis; chronic cystitis, 74 (31.35%), including chronic cystitis un-complicated type, lymphocytic cystitis, plasmolymphocytic cystitis, follicular cystitis, hyperplasia, nodular/acinar hyperplasia, and cystolithiasis; and nothing unusual diagnosed, 118 (50.00%). Similar types of pathological conditions were diagnosed in both species of animals with exception of follicular cystitis and nodular/acinar hyperplasia which was diagnosed respectively only in buffaloes and cystoliths in cows. In addition, a good number of 17/25 (68%) urinary bladder samples tested were found positive for presence of bovine papillomavirus type-2 (BPV-2) by polymerase chain reaction. These included eight cases of acute cystitis, an equal number of cases of chronic cystitis, and one normal bladder. BPV-2 is known as potential source of enzootic bovine hematuria along with other co-factors in enzootic areas. Lesions of zoonotic significance, like tuberculosis, etc., were not diagnosed. None of the observed lesions represented conditions, which, by themselves, would warrant carcass condemnation in buffaloes.

Preliminary assessment of binary ethylenimine inactivated and saponized cutaneous warts (BPV-2) therapeutic vaccine for enzootic bovine haematuria in hill cows.

A preliminary therapeutic vaccine trial was conducted in hill cows to evaluate the therapeutic potential of binary ethylenimine (BEI) inactivated and saponized bovine papillomavirus-2 (BPV-2) for enzootic bovine haematuria (EBH). Although the vaccine failed to show favorable clinical vaccine results in treatment of EBH affected cows at 120 days post-vaccination but immunopathological responses were encouraging. A significant difference was observed in humoral (against Brucella abortus strain 19S) and cell-mediated (in vivo phytohaemagglutination delayed type hypersensitivity (PHA DTH) test and CD4+/CD8+ T-cells ratio by FACS analysis) immune responses following vaccination. The vaccinated animals grossly failed to show regression of bladder tumours but microscopically engorgement and marked perivascular infiltration of mononuclear cells was observed which are indicative of the induction of initial stages of tumour regression. Overall results indicated that the therapeutic vaccine developed can have potentials for treating EBH in cows, for which further modifications in vaccine dose and field trial is required.

Cystosonography as an aid in diagnosis of urinary bladder neoplasms in enzootic bovine haematuria

To ascertain the status of urinary bladder tumors (UBTs) in clinical cases of Enzootic Bovine Haematuria (EBH), the cystosonographic examination was evaluated as an aid in diagnosis. The urinary bladder of EBH affected animals showed thick bladder wall along with presence of some space occupying lesions suggestive of tumors. It also showed anechoic images indicating distended urinary bladder. The ecogenicity of urine content was not homogenous in some cases revealing swirling material. These findings were correlated with urinalysis which revealed that EBH affected animals were either microhaematuric or macrohaematuric. On necropsy, multiple hanging cauliflower-like growths or nodular thickenings in urinary bladders in EBH affected animals were observed. Histopathologically, majority of UBTs were diagnosed as transitional cell carcinoma-papillary type. Post-sonographic urinalysis, necropsy and histopathological diagnosis of these cases confirmed sonographic findings. This technique was found simple, accurate and reliable and can be applied in diagnosis and prognosis of UBTs in EBH cases.