Molecular detection of enterotoxins in multidrug resistant Aeromonas from ready to eat foods in North Western Himalayas: Public health significance.

Aeromonas spp. are normal inhabitants of aquatic environments and are emerging foodborne bacterial pathogens. Aeromonas spp. contamination is frequent in ready-to-eat (RTE) seafood and can also occur in products prepared from milk or meat. The study determined the enterotoxin and antimicrobial susceptibility profiles of Aeromonas spp. isolates recovered from RTE milk products (n = 105), RTE meat/fish products (n = 40) and drinking water (n = 60) samples collected from tourist places in Himachal Pradesh, India, in northwestern Himalayas. 7.3 % (16/220) samples were found contaminated with Aeromonas spp. These isolates were identified as A. hydrophila (31.3 %), A. schubertii (25.0 %), A. sobria (25.0 %) and A. veronii (18.8 %). Aeromonas spp. contamination was significantly higher (14.3 %, 15/105, p = 0.0001) in RTE milk products. The contamination levels for water samples were 1.7 % whereas none of the tested RTE meat or fish products yielded Aeromonas spp. Among RTE milk products, contamination was significantly higher in paneer (South Asian soft cheese) (26.1 %, p = 0.0027) and cream (25.0 %, p = 0.046) based RTE foods. All isolates carried alt (361 bp), encoding a cytotonic heat-labile enterotoxin. Ampicillin resistance was 100 % and high levels (>30 %) of resistance were recorded for amoxicillin/clavulanic acid, amikacin, cefotaxime and ceftazidime. Six (37.5 %) isolates were multi drug resistant (MDR), showing resistance to aminoglycosides, cephams and penicillins. Isolation of alt carrying MDR isolates from RTE foods indicates that Aeromonas spp. can be potential foodborne public health threat in northwestern Himalayas.

Diagnosis of rabies using reverse-transcription polymerase chain reaction on post-mortem skin tissue specimens of the nasolabial plate in a rabies suspected cow: a case study.

In India, rabies in cattle is under-reported. Religious sentiments hamper its diagnosis, discouraging post-mortem examination, particularly opening the cranium. Specimens of peripheral tissue innervated by the cranial nerves could potentially be used as alternative diagnostic specimens to the brain. Herein, we present a case study of a novel approach for diagnosing rabies in a cow suspected of having rabies, using skin tissue specimens of the nasolabial plate obtained post-mortem. Brain and nasolabial tissue specimens tested positive for rabies using conventional reverse-transcription polymerase chain reaction. This approach has been previously shown to have a high diagnostic sensitivity in animals. We encourage further studies with more nasolabial plate skin specimens for both post- and antemortem diagnosis of rabies in cattle.

Sweet syndrome unresponsive to corticosteroid therapy managed with ayurveda medication: A case report.

BACKGROUND: Sweet syndrome (SS) is characterized by sudden appearance of multiple, painful erythematous, papular-nodular skin lesions accompanied by a fever and myalgia. SS resembles Visphota (Bullous eruption disorders) in Ayurveda. SS is a scarcerare skin condition, and Ayurvedic management of this condition is not reported. Therefore, this case report is prepared after taking informed consent from the patient. CASE PRESENTATION: A 34-year-old male patient presented with a complaint of the appearance of multiple blisters over the upper and lower extremities, forehead, and chest with mild fever, congestion of the eye, joint pain, and muscular stiffness for the last two weeks. He has been on oral prednisolone for ten weeks by a dermatologist. The patient has no history of taking drugs in the previous four months before the onset of the skin lesions. The patient was prescribed Nagaradi Kasya (Amrttam Kasaya), Siddha Makardwaja (Plain), Kaisara Guggulu (KG), Ashwagandha ghana vati, and Avipatikar Churna in recommended dose for two weeks. Fifty percentage lesions disappear along systemic symptoms within seven days and complete remission of lesions in fifteen days. The medication was stopped after fifteen days and observed for one year. There is a significant reduction of inflammatory markers like-erythrocyte sedimentation rate (ESR), Total Leucocyte Count (TLC), and C Reactive protein (CRP) after the treatment compared to the baseline also. CONCLUSION: Ayurvedic modalities may be a better treatment option in rare skin conditions like sweet syndrome, where corticosteroid usage failed to improve. More evidence of the usefulness of the Ayurvedic therapeutic approach must be gathered.

AYUSH- 64: A potential therapeutic agent in COVID-19.

Corona Virus disease (COVID-19) has become a global pandemic resulting in large scale morbidity and mortality worldwide. The management of COVID-19 has been a big challenge because of multifactorial pathophysiology and no specific treatment. AYUSH-64, a poly-herbal formulation developed by CCRAS, Ministry of AYUSH, Govt. of India through extensive pharmacological, toxicological and clinical studies has proven its safety and efficacy in infective febrile conditions such as malaria and influenza. AYUSH- 64 has four ingredients having immunomodulatory, anti-inflammatory, antipyretic, antioxidant and anti-viral activities. It arrests the extreme inflammatory responses in COVID-19 that causes progression to significant morbidity. AYUSH-64 has also been incorporated in the National COVID management protocol based on Ayurveda and Yoga by Government of India for asymptomatic and mild cases of COVID-19. Further, on the basis of tangible evidence generated through robust clinical and experimental studies on AYUSH-64, the Ministry of AYUSH has launched nation-wide campaign for mass distribution of AYUSH-64 to asymptomatic, mild to moderate COVID-19 patients in home isolation to reduce the burden on the hospital. This review will highlight the specifications of AYUSH-64, its mechanism of action, its repurposing for COVID-19, various clinical and experimental studies.

Determination of antibiotic residues in bovine milk by HPLC-DAD and assessment of human health risks in Northwestern Himalayan region, India.

Antibiotic residues in milk affects economics of dairy industry and poses health risks to consumers. This study aimed to assess health risks associated with presence of antibiotics in 173 raw and pasteurized milk sampled from northwestern Himalayan state of India. The oxytetracycline and amoxicillin were quantitatively analyzed using validated HPLC-DAD. Methods were selective and linear (R2 > 0.99) with decision limit and detection capability of 1.4 and 0.9 µg/kg and 2.5 and 1.5 µg/kg for oxytetracycline and amoxicillin, respectively. Recoveries ranged from 88-98% with relative standard deviation < 10%. Oxytetracycline and amoxicillin were detected in 8.1% and 1.2% samples, with 1.7% and 1.2% samples exceeding the tolerance limits, respectively. Health risk assessment revealed that estimated daily intakes of antibiotics through milk were lower than acceptable daily intakes (ADI). However, children might receive 9-21% of determined ADI through milk consumption only. Therefore, continuous, sub-therapeutic and long term exposures of antibiotics can pose health risk to consumers. Hence, current findings elucidate the need for vigilant monitoring of antibiotics accompanied by educational programs to farmers for adopting good husbandry practices and adherence to withdrawal periods to meet the expectations of food safety and safeguarding human health. SUPPLEMENTARY INFORMATION: The online version of this article contains supplementary material available at (10.1007/s13197-021-04988-8).

Bacillus cereus: public health burden associated with ready-to-eat foods in Himachal Pradesh, India.

The study determined incidence, enterotoxigenecity and antimicrobial susceptibility profiles of Bacillus cereus isolated from ready-to-eat (RTE) milk products (n = 80), RTE meat products (n = 40), beverages (n = 40) and water samples (n = 60, from food preparing and serving outlets/restaurants) collected from eight different tourist places of Himachal Pradesh. 11.4% (25/220) samples were contaminated with Bacillus and isolates were identified as B. cereus (76.0%, n = 19), B. alvei (12.0%, n = 3), B. polymyxa (8.0%, n = 2) and B. firmus (4.0%, n = 1) by conventional and molecular methods. B. cereus incidence was highest in cheese based foods (25.0%) followed by vegetable soups (16.7%), khoa based foods (14.0%), milk based beverages (10.5%), paneer based foods (8.6%), cream based foods (8.3%) and water (8.3%) samples. Multiplex polymerase chain reaction detected enterotoxigenic genes only in B. cereus isolates. nhe complex (encoding non-haemolytic enterotoxins, ABC) genes were detected only in B. cereus isolates. 57.6% (11/19), 36.8% (7/19) and 5.3% (1/19) harboured all three (nheA, nheB, nheC), two (nheB, nheC) and one (nheC) nhe gene, respectively. Among hbl complex genes (encoding haemolytic enterotoxins CAD), only hblC (36.8%, 7/19) was detected. Incidence B. cereus cytK (encoding cytotoxin enterotoxin) was 52.6% (10/19). Each B. cereus isolate harboured two or more enterotoxigenic genes. Seven isolates had at least one gene from haemolytic and non-haemolytic complexes along with cytK. High levels (> 50%) of antimicrobial resistance were recorded for penicillin, amoxicillin, ampicillin cefixime and ceftazidine in tested B. cereus isolates. Two isolates were identified as multidrug resistant isolates with resistance to ≥ 3 antibiotic classes.

Incidence and antimicrobial susceptibility of Staphylococcus aureus isolated from ready-to-eat foods of animal origin from tourist destinations of North-western Himalayas, Himachal Pradesh, India.

This study was aimed to determine the incidence of Staphylococcus aureus in ready-to-eat (RTE) milk (n = 120) and meat (n = 120) products from various tourist places in north western Himalayas, Himachal Pradesh, India. S. aureus isolates and its enterotoxins; A, B, D and E were characterized by conventional and molecular methods. Antimicrobial susceptibility (AMS) profiles of S. aureus isolates were determined by disk diffusion method using Clinical and Laboratory Standards Institute criteria. Overall, 6.7% (n = 16/240) food samples were positive for S. aureus. PCR amplification of nucA confirmed all biochemically characterized isolates as S. aureus. Incidence of S. aureus was higher (10.0%) in RTE milk products than meat products (3.3%). S. aureus contamination levels were highest in milk cake/khoa (26.0%, p = 0.0002) followed by ice cream/kulfi (10.0%, p = 0.4), mutton momo (10.0%, p = 0.4), burfi (3.3%, p = 0.7) and chicken momo (3.3%, p = 0.7). None of the isolates carried genes for S. aureus enterotoxins; A, B, D and E. AMS testing revealed seven different resistance patterns and 81.3% multi drug resistance. All the isolates were resistant to ampicillin. High resistance levels were observed against methicillin (93.7%), clindamycin (68.8%), erythromycin (56.3%) and vancomycin (43.8%). Vancomycin resistant (n = 7) isolates were also resistant to methicillin. All isolates were susceptible to novobiocin.

A single case study of treating hypertrophic lichen planus with Ayurvedic medicine.

Ayurvedic medicines are often considered effective for chronic and lifestyle disorders. Hypertrophic lichen planus (HLP) is a rare inflammatory skin condition and develops into squamous cell carcinoma in few cases. It has resemblance with Charma Kushtha mentioned in Ayurvedic classics. Conventional therapy used in this condition is unsatisfactory and is not free from side effects. A case of long-standing systemic steroid-dependent HLP is presented here which was intervened successfully with Ayurvedic modalities.

The prevalence of dyslipidemia in patients with diabetes mellitus of ayurveda Hospital.

BACKGROUND: Dyslipidemia is one of the major risk factors for cardiovascular disease in diabetes mellitus. Early detection and treatment of dyslipidemia in type-2 diabetes mellitus can prevent risk for atherogenic cardiovascular disorder. The rationale of this study was to detect the lipid abnormality in diabetic patients. METHODS: Necessary data was collected from the medical archives of 150 patients (73 female and 77 male) with diabetes mellitus registered in Department of pathology and biochemistry of a Ayurveda hospital established at Kolkata, India. RESULTS: The mean ages of female and male subjects were 51.8 ± 10.8 and 53.2 ± 11.3 years respectively. The range and mean value of FBS in females were 113-342 mg/dl and 157.7 ± 6.3 mg/dl, while the range and mean value of PPBS in females were 135-560 mg/dl and 275.5 ± 12.3 mg/dl respectively. Results showed that range and mean value of FBS in males were 111-462 mg/dl and 160.8 ± 7.4 mg/dl, while the range and mean value of PPBS in males were 136-598 mg/dl and 302.1 ± 12.6 mg/dl respectively. Results of serum lipids showed that the mean values for total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C) and very low density lipoprotein cholesterol (VLDL-C) in female patients were 202.2 ± 5.9 mg/dl, 168.3 ± 8.2 mg/dl, 44.9 ± 1.3 mg/dl, 123.6 ± 5.2 mg/dl and 33.7 ± 1.7 mg/dl respectively. The mean values for TC, TG, HDL-C, LDL-C and VLDL-C in male patients were 182.5 ± 4.8 mg/dl, 128.1 ± 10.8 mg/dl, 40.8 ± 1.2 mg/dl, 105.4 ± 4.8 mg/dl and 36.2 ± 2.2 respectively. FBS showed significant positive correlation with PPBS, cholesterol, TG, and VLDL-C. PPBS also demonstrated direct and significant correlations with TG and VLDL-C. CONCLUSIONS: The study showed common lipid abnormalities during diabetes induced dyslipidemia i.e., hypercholesterolemia, hypertriglyceridemia and elevated LDL-C. This study suggests the dominance of hyperlipidemia over increased prevalence of dyslipidemia.

The folklore medicinal orchids of Sikkim.

BACKGROUND: Orchids are well-known for decorative and aromatic values than its medicinal properties. Jivanti, Jivaka, Rsabhaka, Rasna, Manakanda, Pañcagula are used in Ayurveda are said to be orchids. There are 50 species of orchids in medicine. Sikkim has identified 523 species of wild orchids so far. AIM: The aim of this study is to determine the folklore medicinal use of orchids in Sikkim. MATERIALS AND METHODS: To assess the traditional medicinal uses of orchid species, close contacts were made with native people particularly, traditional healers, religious leaders, nursery growers and villagers of Sikkim. The information was gathered with the help of the questionnaire and personal interviews with various knowledgeable respondents during the field visit in between August 2009 and December 2011. RESULTS AND CONCLUSION: We found that 36 species of orchids are used as medicines for different purposes of health. The botanical and ayurvedic name, phenology, parts used and medicinal uses of 36 orchids are presented in this paper along with its local distribution.

Microscopic characterization of Scoparia dulcis Linn.(Scrophulariaceae).

This manuscript covers a detailed pharmacognostic evaluation of Scoparia dulcis Linn. whole plant (Scrophulariaceae), including morphology, microscopy, physicochemical, and phytochemical screening. Microscopy of different plant part was done by performing transverse sections and longitudinal sections, which were identified by the different staining reagents and dyes. Physicochemical constants were done for whole plant; it includes ash value, extractive value and moisture content. Phytochemical screening was done for aqueous and methanolic extract in maceration and soxhletion, results revealed the presence of alkaloids, glycosides, carbohydrates, phenolic compound, flavonoids, saponins, proteins, and amino acids. These study includes parameters to establish the authenticity of S. dulcis and can possibly help to differentiate the drug from its other species.

Sero-prevalence of brucellosis in occupationally exposed human beings of Himachal Pradesh (India).

The chief objective of respective study was to investigate the seroprevalence of brucellosis among occupationally exposed human beings in Himachal Pradesh. A total of 165 serum samples that were obtained from human beings from various regions of the state were screened through a battery of serological tests which included RBPT, STAT, 2-MET, dot-ELISA and indirect-ELISA. 165 of human sera samples included 42 from veterinarians, 40 shepherds, 35 livestock owners, 20 workers at veterinary hospitals/clinics, 16 abattoir workers and 12 veterinary pharmacists. The overall seroprevalence of brucellosis among occupationally exposed human beings was observed to be 6.66% showing highest in abattoir workers (18.75%) followed by pharmacists (8.33%), veterinarians (7.14%), and livestock owners (5.71%) and shepherds (5.00%). In humans it is prevalent as an occult infection or under diagnosed disease, especially; in case of abattoir workers the highest seropositivity for brucella agglutinins was observed. Indirect-ELISA and Dot-ELISA proved best in the diagnosis of brucellosis.

Traditional uses and medicinal potential of Cordyceps sinensis of Sikkim.

Cordyceps sinensis has been described as a medicine in old Chinese medical books and Tibetan medicine. It is a rare combination of a caterpillar and a fungus and found at altitudes above 4500m in Sikkim. Traditional healers and local people of North Sikkim recommend the mushroom, i.e., Yarsa gumba, Keera jhar (C. sinensis) for all diseases either as a single drug or combined with other herbs. The present study was undertaken to collect information regarding the traditional uses of cordyceps in Sikkim. It was found that most local folk healers/traditional healers use cordyceps for the treatment of 21 ailments. A modern literature search was carried out to assess whether the curative effects are valid or just blind faith of local people. Chemical constituents of cordyceps are given and pharmacological and biological studies reviewed. More mechanism-based and disease-oriented clinical studies are recommended.

Clinical efficacy of herbal Padmapatradi yoga in bronchial asthma (Tamaka Swasa).

Ayurveda refers to bronchial asthma as Tamaka Swasa and it is well explained in Charaka Samhita. It contributes several modalities of the treatment for Swasa roga(asthma). Among all modalities of treatment, polyherbal combinations are said to be well-accepted, safe and effective in asthma. A study was carried out in 40 patients of either sex in between the age of 15-65 years to assure the clinical response of Padmapatradi yoga in bronchial asthma (Tamaka Swasa) at P.G. department of Kayachikitsa, D.G.M. Ayurvedic Medical College, Gadag, Karnataka. The sum total properties of Padmapatradi yoga is tikta katu rasa, laghu and tikna guna (light and penetrating properties), ushna virya (hot potency) and vatakaphagna (decrease vata and kapha dosa) Padmapatradi yoga is effective in increased peak expiratory flow rate, breath holding time, and reduces the absolute eosinophil count of studied cases and also found statistically highly significant at p<0.001 level. The drug is quite safe and acts as a bronchodilator, antihistaminic and anti-inflammatory.

Puttur kattu (bandage) - A traditional bone setting practice in south India.

Traditional bone setting practices are quite popular in India and nearly 6000 traditional bone setting Vaidyas (Practisioners) are practicing the same in our country. Puttur kattu is a traditional way of bone setting practice, invented accidentally by K. Kesava Raju in 1881. Now, the fourth generation of his family is practicing this bone setting practice in hospitals at Puttur, Andhra Pradesh, with 200-300 patients per day. A prospective study was undertaken to analyze the techniques in diagnosis, way of management, medicine preparation, plants used and way of applications by traditional bone setter (TBS) Vaidyas, with special reference to Puttur. We also tried to understand the reasons which make lots of people go to Puttur for getting treatment, means of contact for treatment, pathology of fracture and outcome of some treated cases through this study. 54% of the studied patients came to Puttur TBS on the advice of old patients. It is observed that more educated people are patronizing this therapy and 23% patients of the observed cases took discharge from modern hospital voluntarily to receive Puttur kattu treatment. 80% patients believed that this therapy with home remedy would fasten the healing process. 44% patients opted for this therapy due to fear of pain, heavy plaster of Paris bandage, prolonged period of immobilization, surgery and amputation. 71% patients of the followed cases were satisfied with the treatment of TBS of Puttur with minimum complications. The authors also attempted to put forth the legacy of the tradition, the way of management and the plant used for bone setting by the Puttur bone setting Vaidyas.

Overdose effect of aconite containing Ayurvedic Medicine ('Mahashankha Vati').

There are chances that the use of larger than recommended dose of Ayurvedic medicines containing aconite can produce drug reactions. Vatsanabha (Aconitum ferox Wall.) is a very well-known ingredient of Ayurvedic formulations and is prescribed as an antipyretic, analgesic, anti-rheumatic, appetizer and digestive. The recommended dose of purified Vatsanabha (A. ferox Wall.) root is 15 mg. We present a case of hypotension and bradycardia due to aconite poisoning caused by overdosing of an Ayurvedic medicine (Mahashankha Vati), which was primarily managed by Ayurvedic treatment.

Mutations in the connexin 29 gene are not a major cause of nonsyndromic hearing impairment in India.

Mutations in the GJC3 gene are known to cause nonsyndromic hearing impairment (NSHI). In this study, we screened for mutations in the connexin 29 (Cx29) gene in peripheral blood collected from patients with NSHI. DNA was extracted from peripheral blood cells of 123 NSHI patients and 127 normal-hearing control subjects. Coding regions of Cx29 were amplified by polymerase chain reaction using primer pairs flanking both exons. Sequences were analyzed and compared with the published Cx29 sequence. On comparison with control subjects, only one patient and her normal-hearing mother showed a novel heterozygous variant in exon 1 c.569T>A (p. Ile190Asn), which most likely represents a rare polymorphism. From the study, we conclude that mutations in the Cx29 gene do not play a role in the causation of NSHI in Indian population.

Health traditions of Sikkim Himalaya.

Ancient medical systems are still prevalent in Sikkim, popularly nurtured by Buddhist groups using the traditional Tibetan pharmacopoeia overlapping with Ayurvedic medicine. Traditional medical practices and their associated cultural values are based round Sikkim's three major communities, Lepcha, Bhutia and Nepalis. In this study, a semi-structured questionnaire was prepared for folk healers covering age and sex, educational qualification, source of knowledge, types of practices, experience and generation of practice, and transformation of knowledge. These were administered to forty-eight folk healers identified in different parts of Sikkim.490 medicinal plants find their habitats in Sikkim because of its large variations in altitude and climate. For 31 commonly used by these folk healers, we present botanical name, family, local name, distribution, and parts used, together with their therapeutic uses, mostly Rheumatoid arthritis, Gout, Gonorrhea, Fever, Viral flu, asthma, Cough and Cold, indigestion, Jaundice etc. A case treated by a folk healer is also recounted. This study indicates that, in the studied area, Sikkim's health traditions and folk practices are declining due to shifts in socio-economic patterns, and unwillingness of the younger generation to adopt folk healing as a profession.

The medico historical perspective of vitiligo (Switra).

The history of Switra is as old as the human civilization. The earliest medical literature has given the description of Switra in rgaveda and Atharvaveda. Switra is also narrated in Mahabharat and in Vinayapitika. A Roman physician known as Celsus first introduced the term Vitiligo in the second century A.D. Vitiligo is also known as Leucoderma. The history of psychosocial repercussion was described in Vedic literature and it is also found in our society. A population-based study was presented to understand the social implication of this disease. The detailed description of etiological factors and modalities of treatment is found in all ayurvedic classics. Bakuci (Psoralia cordifolia) is still continuing as a photosensitizing drug in the treatment of Switra from the time of Caraka.