Pradip Kumar Das

Evaluation of autologous bone marrow in wound healing in animal model: a possible application of autologous stem cells

A study was conducted to evaluate the potential of autologous bone marrow‐derived cells in comparison with buffy coat of autologous blood for rapid cutaneous wound healing in rabbit model. Three square full‐thickness skin excisional wounds were created in 15 selected experimental animals (rabbit) divided randomly into three groups. The wound was treated with autologous bone marrow cells in plasma (group 1), buffy coat of blood in plasma (group 2) and autologous plasma as control (group 3). Wounds were observed for 30 days for granulation tissue formation, biochemical, histomorphological and histochemical evaluation. In this study, granulation tissue appeared significantly lesser in wounds of group 3 animals followed by group 2 and 1 animals. Neovascularisation, granulation tissue formation, denser, thicker and better arranged collagen fibres, reticulin fibres and elastin fibres formation was more in group 1 as compared with other groups. It was concluded that the application of bone marrow‐derived nucleated cells into the wound margins resulted in early and significantly faster rate of complete healing as compared with buffy coat of autologous blood and autologous plasma (control). This approach may be beneficial in various surface wounds that heal at a slower rate and recommended for healing of various complicated wound in future.

Enhanced bone regeneration with carbon nanotube reinforced hydroxyapatite in animal model

In order to improve the inherently poor mechanical properties of hydroxyapatite (HAp) and to increase its feasibility as load bearing implant material, in the present investigation, functionalised (HFC1 and HFC2) and non-functionalized (HC1 and HC2) multi-walled carbon nanotubes were used as reinforcing material with HAp. Significant improvement with respect to fracture toughness, flexural strength and impact strength of the composites was noticed. In vitro biological properties of HAp-carbon nanotube (CNT) biocomposites have also favored uniform and systematic apatite growth on their surface. Subsequently, in vivo osseous ingrowth at bone defect of rabbit femur was evaluated and compared using radiology, push out test, fluorochrome labeling, histology and scanning electron microscopy after 2 and 4 months respectively. The results demonstrated growth of web like soft callus from the host bone towards the implant, ensuring strong host bone interaction. Toxicological studies of the liver and kidney cells exhibited no abnormality, thereby confirming non-toxicity of the CNT in the animal body. Host-implant biomechanical strength showed high interfacial strength of the composites, indicating their high potentials to be used for bone remodeling applications.

Virulence Repertoire, Characterization, and Antibiotic Resistance Pattern Analysis of Escherichia coli Isolated from Backyard Layers and Their Environment in India

SUMMARY This study was undertaken to observe the prevalence, serogroup, avian pathogenic Escherichia coli (APEC)-associated virulence gene, randomly amplified polymorphic DNA (RAPD) pattern, and antibiotic resistance genes of E. coli in backyard layers and their environment in India. From the 360 samples of healthy layers and their environment, 272 (75.5%) E. coli were isolated. The majority (28.67%) of them were untypeable. Among the studied virulence genes (papC, tsh, iucC, astA), 52 (14.32%) isolates were found to possess astA, including the isolates from the drinking water of the birds (4/272, 1.47%). These strains belonged to 18 different serogroups. Most of the isolates were typeable by RAPD and they produced different patterns. Phenotypic resistance of the isolates was most frequently observed to erythromycin (95.83%), chloramphenicol (87.52%), and cotrimoxazole (78.26%). None of the isolates was found to possess extended-spectrum beta-lactamases (blaTEM, blaSHV, blaCTX-M) or quinolone resistance (qnrA) genes by PCR. The present study was the first attempt in India to assess APEC distribution in backyard poultry production. RESUMEN Repertorio de virulencia, caracterización y análisis de los patrones de resistencia a los antibióticos de Escherichia coli aisladas de gallinas de postura de traspatio y de su medio ambiente en la India. Este estudio se realizó para determinar la prevalencia, el serogrupo, los genes asociados a virulencia de Escherichia coli patógena para las aves (APEC), los patrones de ADN polimórfico amplificado aleatoriamente (RAPD), y los genes de resistencia a los antibióticos de E. coli en gallinas de traspatio y su ambiente en la India. De las 360 muestras de gallinas sanas y de su ambiente, se aislaron 272 cepas de E. coli (75.5%). La mayoría (28.67%) de ellas no fueron tipificables. Entre los genes de virulencia estudiados (papC, tsh, iucC, astA), se encontró que 52 aislamientos (14.32%) poseían el gene astA, incluyendo los aislamientos de agua de bebida de las aves (4/272, 1.47%). Estas cepas pertenecían a 18 serogrupos diferentes. La mayoría de los aislamientos fueron tipificables mediante RAPD y produjeron diferentes patrones. La resistencia fenotípica de los aislamientos se observó con mayor frecuencia contra eritromicina (95.83%), cloranfenicol (87.52%), y contra cotrimoxazol (78.26%). Se encontró mediante PCR que ninguno de los aislamientos poseía genes de un espectro extendido de beta-lactamasas (blaTEM, blaSHV, blaCTX-M), o resistencia contra quinolonas (qnrA). El presente estudio es el primer intento en la India para evaluar la distribución de E. coli patógena para la producción de aves de traspatio.

A comparative study on the expression profile of MCTs and HSPs in Ghungroo and Large White Yorkshire breeds of pigs during different seasons.

Thermal stress has a significant adverse effect on commercial swine production but it is not easy to measure. Animals may adapt to stress conditions by an alteration in the expression of stress-related genes such as heat shock proteins (HSPs) and monocarboxylate transporters (MCTs). The present study presents a comparative analysis of seasonally varied effects on the expression profiles of HSPs (27, 70, and 90) and MCTs (1, 2, and 4) transcripts in thigh muscle and colon tissue of Ghungroo and Large White Yorkshire (LWY) breeds of pig. By real-time polymerase chain reaction, the mRNA expression of HSP27 and HSP90 genes was found to be higher in both thigh muscle and colon tissue in Ghungroo compared to Large White Yorkshire pigs during the summer. However, the relative expression of HSP70 was significantly higher (P < 0.01) in Ghungroo compared to Large White Yorkshire pigs during both seasons in both thigh muscle and colon tissue. The expression of HSP90 was higher in Ghungroo when compared to LWY though the variation was non-significant (P > 0.05) in the colon during different seasons. However, in Ghungroo, the mRNA expression of MCT1 was found to be significantly (P < 0.05) higher in thigh muscle and colon regions during the summer compared to LWY, whereas MCT2 was expressed more in the colon in LWY compared to Ghungroo during the summer. The relative expression of mRNA of MCT4 was found to be significantly (P < 0.05) higher in thigh region in both summer and winter in Ghungroo compared with LWY. Thus, the study demonstrated that both HSPs and MCTs gene expression during thermal stress suggests the possible involvement of these genes in reducing the deleterious effect of thermal stress, thus maintaining cellular integrity and homeostasis in pigs. These genes could be used as suitable markers for the assessment of stress in pigs.

Electrocardiogram of Clinically Healthy Mithun (Bos frontalis): Variation among Strains

A study was conducted to establish the normal electrocardiogram in four different genetic strains of mithun (Bos frontalis). Electrocardiography, cardiac electrical axis, heart rate, rectal temperature and respiration rate were recorded in a total of 32 adult male mithun of four strains (n = 8 each). It was found that the respiration and heart rates were higher (P < .05) in Manipur than other three strains. Amplitude (P < .05) and duration of P wave and QRS complex differed (P < .01) among the strains. Mizoram strain had the highest amplitude and duration of P wave and QRS complex. On the other hand, higher (P < .05) amplitude and duration of T wave were recorded in Arunachalee and Mizoram strains. The mean electrical axis of QRS complex that were recorded for Arunachalee and Manipur strains were similar to that reported for other bovine species; whereas the electrical axis of QRS for Nagamese and Mizoram strains were more close to feline and caprine species, respectively. In conclusion, electrocardiogram of mithun revealed that the amplitude and duration of P wave, QRS complex and T wave were different among four different genetic strains of mithun and the electrical axis of QRS complex for Nagamese and Mizoram mithuns are dissimilar to bovine species.

Alteration in the in vitro activity of milk leukocytes during different parity in high yielding cross-bred cows

Abstract In vitro activity of milk leukocytes (viz. neutrophils, lymphocytes and macrophages) was evaluated in forty-eight (48) clinically healthy high-yielding cross-bred cows of mid-lactation stage (100–200 days of lactation), divided into four groups namely 1st parity (n = 12), 2nd parity (n = 12), 3rd parity (n = 12) and 4th and above parity (n = 12). Milk samples were taken (250 ml/cow) were taken. Milk somatic cell counts (SCC) and differential leukocyte counts (DLC) were performed microscopically. In vitro phagocytic index (PI) of milk neutrophils and macrophages was evaluated by colorimetric nitro blue tetrazolium reductive assay. Mitogen-induced milk lymphocyte blastogenic response was measured by colorimetric MTT (tetrazolium) assay after isolation of the milk leukocytes by density gradient centrifugation. Milk SCC differed significantly (p < 0.01) between different parity. Cows of 4 and above parity showed significantly (p < 0.01) higher milk SCC compared to primiparous cows. There was no significant difference in milk DLC during different parities in high-yielding cross-bred cows. There was a significant (p < 0.01) variation in lymphocyte blastogenesis amongst parity. The highest value of lymphocyte blastogenesis was seen at 3rd parity, whereas lowest value was obtained in the cows of both 1st and 4th or above parity. PI of milk neutrophils did not differ significantly between parity. PI of milk macrophages was significantly (p < 0.01) higher in 3rd parity and lower (p < 0.01) in 1st and 4th parities. The study indicated that depressed activity of milk lymphocytes and macropages was lower and SCC was higher in the cows of 4th and above parity indicating more mammary stress and hence susceptible to udder infection and mastitis. Therefore, better care and managemental interventions should be taken around these periods.

Association between udder morphology and in vitro activity of milk leukocytes in high yielding crossbred cows

Aim: The present investigation was conducted to study the association between udder morphology and in vitro activity of milk leukocytes in high yielding crossbred cows. Materials and Methods: A total of 48 healthy high yielding crossbred cows were selected for the study. The udder configuration and teat/udder morphology were recorded before milking. Milk samples (100 ml/cow) were collected aseptically. Milk somatic cell counts (SCC) and milk differential leukocyte counts were performed microscopically. Milk leukocytes (viz., neutrophils, lymphocytes, and macrophages) were isolated from milk samples by density gradient centrifugation. Phagocytic index (PI) of milk neutrophils and macrophages were evaluated by colorimetric nitro blue tetrazolium assay. Lymphocytes proliferation response was estimated by MTT assay and expressed as stimulation index. Results: There was a significant (p<0.01) positive correlation between milk SCC with mid teat diameter, teat base diameter and significant (p<0.05) negative correlation between milk SCC and the height of the teat from the ground. Milk SCC was found to be significantly (p<0.01) lower in bowl-shaped udder and higher (p<0.01) in pendulous type. Milk macrophage percentage was positively (p<0.01) correlated with udder circumference. PI of milk neutrophil was negatively (p<0.01) correlation between teat base diameter, and PI of milk macrophages was found to be positively (p<0.01) correlated with teat apex diameter. Both PI of milk neutrophils and macrophages was found to be significantly (p<0.01) lower in the animals having flat and round teat and pendulous type of udder. In vitro PI of milk neutrophils was found to be significantly (p<0.01) lower in flat teat. In vitro PI of milk macrophages was found to be significantly (p<0.01) lower in the round and flat teats compared to pointed and cylindrical teats. Conclusion: Udder risk factors such as teat shape and size, teat to floor distance, udder shape, and size may decrease the in vitro activity of milk leukocytes hence facilitates the incidence intramammary infections.

Electrocardiogram pattern of some exotic breeds of trained dogs: a variation study.

Aim: The present study has been conducted to evaluate the variation in electrocardiogram (ECG) parameters among different trained breeds of dogs (viz. Labrador, German Shepherd, and Golden Retriever) used for security reasons. Materials and Methods: The ECG was recorded by single channel ECG at a paper speed of 25 mm/s and calibration of 10 mm=1 mV. The recordings were taken from all the standard bipolar limb leads (Lead-I, II, and III) and unipolar augmented limb leads (Lead-aVR, aVL, and aVF). Results: Heart rate was found to be highest in Labrador and lowest in German Shepherd. P-wave duration was maximum in Golden Retriever breed and lowest in Labrador. Maximum amplitude of P-wave was found in Labrador followed by German Shepherd and Golden Retriever. There was significantly (p<0.05) higher values of PR interval in German Shepherd compared to other breeds. The variation in QRS duration, ST segment duration, T-wave duration, and T-wave amplitude was found to be non-significant among breeds. Inverted T-waves were most common in Golden Retriever and German Shepherd, whereas positive T-waves were found in Labrador. There was significant (p<0.05) variation in mean electrical axis of QRS complex among different breeds and it ranges from +60° to +80°. Conclusion: The present study provides the reference values for different ECG parameters to monitor the cardiac health status among Labrador, German Shepherd, and Golden Retriever breeds.

Evaluation of egg production after adoption of biosecurity strategies by backyard poultry farmers in West Bengal

Aim: On the basis of identified source of major bacterial infections at four agro-climatic zones in West Bengal the cost-effective biosecurity strategy was formulated for backyard poultry farmers. The aim of the present study was to assess the adoption. So, the study was aimed to detect the adoption level of the formulated biosecurity strategy to mitigate the Salmonella and Escherichia coliweek post-hatch period chicks were contamination level in the sources and its correlation with egg production in West Bengal. Materials and Methods: A questionnaire was prepared querying regarding the biosecurity measures presently followed by the farmers, if any and egg production of their birds. Subsequent to the interview the formulated biosecurity strategy was conveyed. After 3 months, the interview with the same questionnaire was conducted to the same farmers to detect their adoption level. Results: The change in practices were noted in certain parameters which differs significantly (p<0.01 or p<0.05). As a consequence, the average egg production/flock was increased in 3 months after adoption of the strategy (618.2±37.77/flock) in comparison to last 3 months average before adoption of the strategy (495.3±30.00/flock) which also differs significantly (p<0.01). Conclusion: The present study detected the implementation of the biosecurity strategy in backyard poultry farming in West Bengal can substantially benefit the farmers in terms of increased egg production.

Biosecurity Strategies for Backyard Poultry: A Controlled Way for Safe Food Production

Abstract The people of rural and periurban India depend on backyard farming system for poultry meat and eggs. It is used by weaker sections of society, such as marginal farmers as insurance against crop failure, for ready cash and to ensure basic economic returns and the empowerment of women and children. However, backyard farming does not appear to be a promising strategy to achieve the poverty reduction until the production level is increased. The major constraint in increasing the production in backyard birds is microbial infection due to lack of biosecurity knowledge among the farmers. The mitigation of the microbial infection depends on the detection of the infection route and its prevention. The source of microbial infection (Salmonella, Escherichia coli) and their dissemination through the eggs in backyard flocks kept in different agroclimatic zones of West Bengal, a major egg producing state in India, was identified. The pattern of virulence gene specially associated with urinary tract infection and antibiotic resistance genes, such as extended spectrum β lactamase (ESBL) genes of the isolates was studied. The chapter will elaborate the backyard farming including breeds reared, housing, feeding with special emphasis on suggested biosecurity strategies and consequence of the adapted strategy.