Resumo O estudo teve como objetivo analisar os efeitos dos antibióticos promotores de crescimento (AGPs), di-hidrato de oxitetraciclina e fosfato de tilosina na microflora intestinal de frangos de corte. Os AGPs foram fornecidos em diferentes concentrações isoladamente ou em combinações por 42 dias de criação. Amostras fecais foram coletadas do intestino (duodeno, jejuno e ceco) de frangos de corte no 14º, 28º e 42º dias de ensaio. As amostras foram cultivadas em diferentes meios seletivos e a identificação bacteriana foi realizada por diferentes ferramentas de diagnóstico bioquímico e molecular. Os resultados mostraram um efeito significativo dos AGPs no crescimento de microrganismos patogênicos como Escherichia coli e Clostridium perfringens no intestino. Curiosamente, um crescimento prejudicado foi observado para ambas as bactérias, mostrando um efeito significativo (P <0,05) de AGPs em E. coli e C. perfringens nos dias 14, 28 e 42. Este efeito foi observado apenas e em combinação com o uso de AGPs. Os dados mostraram ainda que o efeito foi mais proeminente em combinação e com um aumento da concentração de AGPs. Nenhum comprometimento foi observado no crescimento de L. reuteri em diferentes locais do intestino e duração (14º, 28º e 42º dias). Os resultados mostraram que o uso de AGPs na dieta não tem efeito nocivo nas bactérias benéficas, no entanto, foi observado um crescimento prejudicado nas bactérias nocivas. Sugere-se que uma combinação de AGPs (OXY-1.0+TP-0.5) seja econômica e não tenha efeito prejudicial sobre o frango de corte. O uso de AGPs em uma dose recomendada e por um período de tempo específico é seguro para uso em aves tanto como promotor de crescimento quanto para prevenção de doenças.

Abstract The study was aimed to analyse the effects of antibiotic growth promoters (AGPs), Oxytetracycline di-hydrate and Tylosin phosphate on the intestinal microflora in broiler chicken. The AGPs were provided in different concentrations solely or in combinations for 42 days of rearing. Faecal samples were collected from the intestine (duodenum, jejunum and caeca) of broiler chicken on 14th, 28th and 42nd days of trial. Samples were cultured on different selective medium and bacterial identification was performed by different biochemical and molecular diagnostic tools. Results showed a significant effect of AGPs on the growth of pathogenic microorganisms such as Escherichia coli and Clostridium perfringens in the intestine. Interestingly, an impaired growth was observed for both bacterium showing a significant effect (P<0.05) of AGPs on E. coli and C. perfringens on day 14th, 28th, and 42nd. This effect was observed solely and in combination while using AGPs. Data further showed that the effect was more prominent in combination and with an increase concentration of AGPs. Remarkably, no impairment was seen on the growth of L. reuteri at different sites of intestine and duration (14th, 28th, and 42nd days). The results showed that the use of AGPs in diet has no harmful effect on beneficial bacteria, however, an impaired growth was seen on the harmful bacteria. It is suggested that a combination of AGPs (OXY-1.0+TP-0.5) is economical and have no harmful effect on the broiler chicken. The use of AGPs in a recommended dose and for a specific period of time are safe to use in poultry both as growth promoter and for the prevention of diseases.

Resumo O zinco é um micronutriente essencial necessário para o crescimento ideal das plantas. Ele está presente no solo em formas insolúveis. A solubilização bacteriana da forma indisponível de Zn no solo para a forma disponível é uma abordagem emergente para aliviar a deficiência de Zn em plantas e seres humanos. Bactérias solubilizadoras de zinco (ZSB) podem ser um substituto para fertilizantes químicos de Zn. O presente estudo teve como objetivo isolar e caracterizar espécies bacterianas de solo contaminado e avaliar seu potencial de solubilização de Zn. Bactérias resistentes ao Zn foram isoladas e avaliadas quanto ao seu MIC contra o Zn. Entre as 13 cepas bacterianas isoladas, ZSB13 apresentou valor máximo de MIC de até 30 mM/L. A cepa bacteriana com maior resistência ao Zn foi selecionada para análise posterior. A caracterização molecular de ZSB13 foi realizada por amplificação do gene 16S rRNA que o confirmou como Pseudomonas oleovorans. A solubilização do Zn foi determinada através de ensaio em placa e meio caldo. Quatro sais insolúveis (óxido de zinco (ZnO), carbonato de zinco (ZnCO3), sulfito de zinco (ZnS) e fosfato de zinco (Zn3 (PO4) 2) foram usados para o ensaio de solubilização. Nossos resultados mostram uma zona de halo clara de 11 mm em placas de ágar corrigidas com ZnO. Da mesma forma, ZSB13 mostrou liberação significativa de Zn em caldo alterado com ZnCO3 (17 e 16,8 ppm) e ZnO (18,2 ppm). Além disso, os genes de resistência ao Zn czcD também foram enriquecidos em ZSB13. Em nosso estudo, a cepa bacteriana compreendendo potencial de solubilização de Zn foi isolada e poderia ser usada posteriormente para o aumento do crescimento de safras.

Abstract Zinc is an essential micronutrient that is required for optimum plant growth. It is present in soil in insoluble forms. Bacterial solubilization of soil unavailable form of Zn into available form, is an emerging approach to alleviate the Zn deficiency for plants and human beings. Zinc solubilizing bacteria (ZSB) could be a substitute for chemical Zn fertilizer. The present study aimed to isolate and characterize bacterial species from the contaminated soil and evaluate their Zn solubilizing potential. Zn resistant bacteria were isolated and evaluated for their MIC against Zn. Among the 13 isolated bacterial strains ZSB13 showed maximum MIC value upto 30mM/L. The bacterial strain with the highest resistance against Zn was selected for further analysis. Molecular characterization of ZSB13 was performed by 16S rRNA gene amplification which confirmed it as Pseudomonas oleovorans. Zn solubilization was determined through plate assay and broth medium. Four insoluble salts (zinc oxide (ZnO), zinc carbonate (ZnCO3), zinc sulphite (ZnS) and zinc phosphate (Zn3(PO4)2) were used for solubilization assay. Our results shows 11 mm clear halo zone on agar plates amended with ZnO. Likewise, ZSB13 showed significant release of Zn in broth amended with ZnCO3 (17 and 16.8 ppm) and ZnO (18.2 ppm). Furthermore, Zn resistance genes czcD was also enriched in ZSB13. In our study, bacterial strain comprising Zn solubilization potential has been isolated that could be further used for the growth enhancement of crops.

The purpose of this study was to determine the comparative efficacy of locally-produced yeast to commercial yeast on growth performance, serum biomarkers, and gut histology. Thirty pre-weaned male Lohi lambs were divided into; 1) C (Control; starter diet with no supplement), 2) CY (starter diet with ewe milk supplemented with 1 g/animal/day commercial yeast), and 3) LY (starter diet supplemented with 1 g/animal/day local yeast). Animals were housed individually and fed a starter diet and orts were weighed daily. Dry matter intake (DMI) was determined daily while the body weights were determined on a weekly basis. At the end of the trial, blood samples were collected and analysed for blood urea nitrogen (BUN), glucose, β-Hydroxybutyrate (BHBA), and non-esterified fatty acids (NEFA), while ruminal fluid and tissues were collected for rumen fermentation parameters and histomorphometry. The results revealed that the DMI and average daily weight gain (ADWG) were higher in CY and LY compared to C. The feed efficiency was also better in CY and LY lambs compared to C lambs. The BHBA, BUN, total proteins (TP), cholesterol, creatinine, blood glucose, and tissue histomorphometry were similar in LY and CY supplementation. In conclusion, supplementation with LY produced comparable results to the CY.

Resumo A ocorrência contínua de metais pesados é uma das principais causas de poluição ambiental devido aos seus efeitos tóxicos. A contaminação por metais pesados representa um grande risco para todas as formas de vida encontradas no meio ambiente. Em concentrações mínimas, esses metais são altamente reativos e podem se acumular nas cadeias alimentares e na cadeia alimentar, causando grandes perigos às preocupações com a saúde pública. Amostras de solo foram coletadas no esgoto de Paharang, Faisalabad. Bactérias tolerantes ao cádmio foram isoladas da amostra coletada pelo método da placa de ágar. As colônias separadas individuais selecionadas foram avaliadas quanto às suas concentrações inibitórias mínimas contra Cd. A cepa bacteriana isolada GCFSD01 apresentou valores de CIM de 30 mM/L. A colônia bacteriana que apresentou maior resistência contra o Cd foi selecionada para identificação. Após seleção da maior colônia bacteriana resistente ao Cd, coloração de Gram e diferentes testes bioquímicos foram realizados para a caracterização da bactéria isolada. A caracterização molecular do isolado bacteriano GCFSD01 foi realizada por PCR 16S rRNA confirmando a presença de Bacillus cereus. Após a identificação molecular, as condições ótimas de crescimento da cepa bacteriana também foram verificadas. A cepa GCFSD01 apresentou crescimento ótimo em pH 7 e temperatura de 37 °C. Nosso resultado revelou que a cepa de B. cereus GCFSD01 reduziu 61,3% de Cd após 48 horas. A tolerância a múltiplos metais e a redução de Cd por B. cereus indicam seu potencial para uso posterior na descontaminação do solo poluído.

Abstract Continuous occurrence of heavy metals is a major cause of environmental pollution due to its toxic effects. At minimum concentrations, these metals are highly reactive and can gather in the food chains and food web, causing major dangers to public health concerns. Soil samples were collected from Paharang drain, Faisalabad. Cadmium tolerant bacteria were isolated and evaluated for their MIC against Cd. The isolated bacterial strain GCFSD01 showed MIC value upto 30 mM/L. The bacterial strain with the highest resistance against Cd was selected for further study. Molecular characterization of bacterial isolate GCFSD01 was performed by 16S rRNA which confirmed it as Bacillus cereus. Optimum growth conditions of bacterial strain were also evaluated. Strain GCFSD01 showed optimum growth at pH 7 and 37 °C temperature. Our result revealed that B. cereus strain GCFSD01 reduced 61.3% Cd after 48 hrs. Multiple metal tolerance and Cd reduction by B. cereus indicate its potential for further use for decontamination of polluted soil.

The study evaluated the efficacy of ethanolic extracts of Catharanthus roseus (C. roseus) in altering growth performance, viscera development and intestinal microarchitecture in broilers. Day-old broiler chicks (n = 175) were randomly divided into seven groups (n = 25/group) with 5 replicates (n = 5). The chicks in the control groups were fed a corn-soya-based basal diet, whereas groups 2 - 4 had the same basal diets supplemented with 0.05%, 0.1%, and 0.2% ethanolic leaf extracts (ELE), respectively. Chicks in groups 5 - 7 were fed the basal diet supplemented with 0.05%, 0.1%, and 0.2% ethanolic root extracts (ERE) of C. roseus, respectively. The growth performance and feed conversion ratio remained unchanged, but feed consumption of birds supplemented with 0.1% ERE increased in weeks 1 and 3. The relative length of the small intestine was lower in the 0.05% ERE group compared with the control. Villus height, width, surface area, and lamina propria thickness were higher in the 0.1% ERE group in all three segments of the small intestine than in the control. Broilers supplemented with 0.1% ERE performed better in terms of feed consumption and improved intestinal microarchitecture of the small intestine.

This study was conducted to reveal the dynamics of haematological indicators in pregnant and nonpregnant dairy cows. Sixty multiparous healthy dairy cows were divided into four groups based on the length of time they had been pregnant, namely first, second and third trimesters, and non-pregnant (n=15 each). Blood was collected from each animal, and serum was harvested and stored at -20 °C for biochemical profiling. Concentrations of serum total oxidants (TOC), ceruloplasmin oxidase (CpO) and triiodothyronine (T3) were higher (P <0.05) during the third trimester compared with non-pregnant cows. Serum arylesterase (Ary) concentration was lower (P <0.05) during the second and the third trimesters compared with the non-pregnant cows. The concentration of serum total homocysteine (tHcy) was higher (p <0.05) the third trimester compared with the first and the second trimesters. The concentrations of serum total antioxidants (TAC), paraoxonase 1 (PON1), thyroxine (T4), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were not different in the pregnant and the non-pregnant cows. Thus, TOC, Ary, CpO, tHcy and T3 could be taken as biological markers to assess the progression of pregnancy and to develop management tools to improve health status during late gestation in dairy cows.

The goal of this study was to compare the effects of Bacillus subtilis and the antibiotic enrofloxacin on growth performance, immune response, intestinal histomorphometry and cecal Salmonella numbers in broilers challenged with S. gallinarum. Salmonella-free chicks (n = 240) were allocated to four groups, comprising six replicates (n = 10) each. Group NN was not infected and received no treatment. Group SN was infected with Salmonella and received no treatment. Group SE was infected with Salmonella and treated with the antibiotic enrofloxacin. Group SP was infected with Salmonella and treated orally with the probiotic B. subtilis. Initially the groups that were infected with Salmonella had reduced growth performance compared with NN. By the fourth week, SE and SP had recovered and weighed as much as or more than NN. The SP group had greater cellular immunity and relative weights of the bursa and thymus than the other groups. SN had the lowest villus height in all the segments of the small intestine and a larger Salmonella population in the ceca. Probiotic supplementation reduced the pathological outcomes more than enrofloxacin. Prophylactic use of B. subtilis-type probiotics had similar effects to the use of antibiotics, alleviated the stress related to infection of S. gallinarum, and improved the growth performance, immune function, and gut mucosal development in broilers.

ABSTRACT Although in-feed antibiotics work for broiler chicken´s growth, they are a source of public health hazard. Therefore, there is a need for alternates which can act as growth promoter without deleterious effects on the health of meat consumers. Moringa oleifera is one such phytobiotic which is reported to possess antimicrobial and immuno-modulatory properties. This study investigated the effect of Moringa oleifera leaf powder (MOLP) supplementation on meat quality and bone morphometry of broiler. One-day-old chicks (n=100), divided into five groups (four replicates with n=5), were fed a basal diet (control group) or same diet supplemented with 6, 9, 12 or 15 g/kg MOLP. On d-35, two birds per replicate were euthanized to collect samples of breast muscle, blood and tibia bone. The MOLP supplementation significantly increased pH of breast muscle and ash percentage of tibia bone. The diameter of breast muscle fibres and also weight and weight length index of tibia bone significantly increased with 12 and 15 g/kg MOLP. The water holding capacity (WHC) of breast muscle was significantly higher with 9 and 15 g/kg MOLP; whereas robusticity index of tibia bone significantly decreased with 12 and 15 g/kg MOLP inclusion compared to the control group. In conclusion, dietary supplementation of Moringa oleifera leaf powder (12g/kg) increased pH, water holding capacity, and muscle fibre diameter of breast muscle and also weight, ash percentage and density indices of tibia bone in broiler chickens.

The dietary supplementation of trace minerals and probiotics has long been practiced to reduce the harmful effects of high environmental temperature in poultry. The study aimed to assess the effect of single or combined supplementation of ZnSO4●H2O and probiotic (Protexin®) on the growth performance, gut biometry and immune organs histomorphology in broilers reared under cyclic heat stress. For this purpose, 280 day-old broiler chicks were randomly divided into seven groups designated as control (basal diet), HS (heat stress + basal diet), HS-Zn30 (HS + 30 mg ZnSO4●H2O/kg feed), HS-Zn60 (HS + 60 mg ZnSO4●H2O/kg feed), HS-Pro (HS + 0.1 g Protexin®/kg feed), HS-Zn30-Pro (HS + 30 mg ZnSO4●H2O/kg feed + 0.1 g Protexin®/kg feed), and HS-Zn60-Pro (HS + 60 mg ZnSO4●H2O/kg feed + 0.1 g Protexin®/kg feed). Each group consisted of five replicates (8 birds per replicate). The birds were fed on starter (0 - 21days) and grower (22 - 42 days) diets. From day 22 onwards, the birds were exposed to cyclic heat stress (temperature and relative humidity at 35 ± 1 °C and 75 ± 5% respectively) for 8 hours. The results revealed that birds exposed to heat stress showed significant decrease in body weight (BW) and increase in feed conversion ratio (FCR) compared to the control group. The dietary supplementation of ZnSO4●H2O and probiotic either singly or in combination improved BW and FCR throughout the experimental period in heat-stressed broilers. Area of lymphatic nodule and lymphoid follicle in caecal tonsils and bursa of Fabricius increased in HS-Pro, HS-Zn30-Pro and HS-Zn60-Pro groups compared to HS group. In conclusion, zinc and probiotics combination partially improved performance and immune organs histomorphology in heat-stressed broilers.

The present research aimed to investigate the effects of Bacillus subtilis (B. subtilis) on performance, immune system, gut, and lymphoid organs' microarchitecture in broilers. A total of 120 day-old broiler chicks were randomly distributed into four groups. The birds were fed a corn-soy-based basal diet (BD) (control) or the same BD supplemented with 10% zinc bacitracin (ZnB), 0.05 g/kg or 0.1 g/kg of B. subtilis (BS). The broilers fed 0.1 g/kg of B. subtilis had superior mean bodyweight and lower feed conversion ratio compared with the non-supplemented or ZnB-fed groups. The BS-0.1 group registered higher antibody titer against the Newcastle disease (ND) virus. Cell-mediated immune response post Phytohaemagglutinin-P (PHA-P) injection was attained by both BS-0.1 and BS-0.05 groups. Histomorphological study revealed increased thymus cortical width, and cortex/medulla ratio in BS-0.1 group compared with control. Area of bursal follicles and germinal centres of spleen also improved in BS-0.1 group. Compared to ZnB and control, higher villus height (VH) and villus crypt ratio of the duodenum and jejunum were recorded on day 21, and higher VH of duodenum and ileum was noted on day 35 in BS-0.1 and BS-0.05 groups. In conclusion, B. subtilis-type probiotics contributed positively to better growth performance, improved immune system and modulated morphology of lymphoid organs and gut mucosa in broilers.

This study aimed to investigate the influence of ß-galacto-oligosaccharides (ß-GOS) on growth performance, organ development and intestinal microarchitecture of broilers during heat stress. Day-old chicks (n = 125) were divided into five groups. The control or thermoneutral zone group (TNZ) was raised under standard management until the 35th day. Four groups were exposed to cyclic heat stress (35 °C 8 h/d) from the 22nd to the 35th day. The TNZ and heat stress control (HSCT) groups were fed a corn-based diet and HS + 0.1% ß-GOS; HS + 0.2% ß-GOS; and HS + 0.5% ß-GOS were fed a corn-based diet supplemented with ß-GOS (0.1%, 0.2%, and 0.5%), respectively. Exposure to heat stress reduced feed consumption, feed efficiency and the relative weight of the liver, bursa of Fabricius, and small intestine, compared with the TNZ group. Morphometric evaluation of the small intestine revealed reduced villus surface area, villus height to crypt depth ratio (VH : CD) and intraepithelial lymphocytes (IELs) in all segments, and reduced acidic goblet cells (AGCs) in the ileum of the HSCT group compared with the TNZ group. Compared with the HSCT group, dietary ß-GOS (0.2% and 0.5%) improved the feed efficiency and relative weight of the small intestine. Furthermore, dietary ß-GOS (0.1%) increased villus surface area in the duodenum and IEL count in the small intestine compared with the HSCT group. Dietary ß-GOS 0.5% increased villi surface area (VSA) in the jejunum and ileum, whereas the IEL count in the small intestine and acidic goblet cells (AGCs) in the jejunum and ileum were reduced compared with the HSCT group. In conclusion, dietary supplementation of ß-GOS (0.2% and 0.5%) improved the growth performance and intestinal microarchitecture of broilers during heat exposure, along with partial immune stimulation.

MTT (3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay is commonly used to validate the viability of metabolically active cells. The study was conducted to examine and validate the MTT test to assess the sperm viability of Nili-Ravi buffalo bulls and compare the efficiency of the test with the supra-vital staining technique (eosin-nigrosine) and hypo-osmotic swelling test. Fresh semen samples from breeding Nili-Ravi buffalo bulls (n = 20) were collected using an artificial vagina. After assessing the quality of semen for normal parameters, the MTT assay was carried out in phosphate buffer saline. Results revealed a high significant correlation (r = 0.995) between the viability of sperm and the rate of reduction of MTT. The other proportions of some semen samples showed a weak relationship between the eosinnigrosine method (r = -0.32), hypo-osmotic swelling test (r = -0.12) and motility (r = -0.08). However, the MTT assay was found to be superior to other tests as it was able to determine those sperm which were more than 90% viable. In conclusion, the MTT assay is a simple, robust test that can be used to select Nili-Ravi buffalo bulls on the basis of sperm quality.