An Immobilized Form of a Blend of Essential Oils Improves the Density of Beneficial Bacteria, in Addition to Suppressing Pathogens in the Gut and Also Improves the Performance of Chicken Breeding.

Antimicrobial growth promoters (AGP) are used in chicken production to suppress pathogens in the gut and improve performance, but such products tend to suppress beneficial bacteria while favoring the development and spread of antimicrobial resistance. A green alternative to AGP with the ability to suppress pathogens, but with an additional ability to spare beneficial gut bacteria and improve breeding performance is urgently required. We investigated the effect of supplementation of a blend of select essential oils (cinnamon oil, carvacrol, and thyme oil, henceforth referred to as EO; at two doses: 200 g/t and 400 g/t feed) exhibiting an ability to spare Lactobacillus while exhibiting strong E. coli inhibition ability under in vitro tests and immobilized in a sunflower oil and calcium alginate matrix, to broiler chickens and compared the effects with those of a probiotic yeast (Y), an AGP virginiamycin (V), and a negative control (C). qPCR analysis of metagenomic DNA from the gut content of experimental chickens indicated a significantly (p < 0.05) lower density of E. coli in the EO groups as compared to other groups. Amplicon sequence data of the gut microbiome indicated that all the additives had specific significant effects (DESeq2) on the gut microbiome, such as enrichment of uncultured Clostridia in the V and Y groups and uncultured Ruminococcaceae in the EO groups, as compared to the control. LEfSe analysis of the sequence data indicated a high abundance of beneficial bacteria Ruminococcaceae in the EO groups, Faecalibacterium in the Y group, and Blautia in the V group. Supplementation of the immobilized EO at the dose rate of 400 g/ton feed improved body weight gain (by 64 g/bird), feed efficiency (by 5 points), and cellular immunity (skin thickness response to phytoheamagglutinin lectin from Phaseolus vulgaris by 58%) significantly (p < 0.05), whereas neither yeast nor virginiamycin showed a significant effect on performance parameters. Expression of genes associated with gut barrier and immunity function such as CLAUDIN1, IL6, IFNG, TLR2A, and NOD1 were significantly higher in the EO groups. This study showed that the encapsulated EO mixture can improve the density of beneficial microbes in the gut significantly, with concomitant suppression of potential pathogens such as E.coli and improved performance and immunity, and hence, has a high potential to be used as an effective alternative to AGP in poultry.

Genetic Diversity, Population Structure and Linkage Disequilibrium Analyses in Tropical Maize Using Genotyping by Sequencing.

Several maize breeding programs in India have developed numerous inbred lines but the lines have not been characterized using high-density molecular markers. Here, we studied the molecular diversity, population structure, and linkage disequilibrium (LD) patterns in a panel of 314 tropical normal corn, two sweet corn, and six popcorn inbred lines developed by 17 research centers in India, and 62 normal corn from the International Maize and Wheat Improvement Center (CIMMYT). The 384 inbred lines were genotyped with 60,227 polymorphic single nucleotide polymorphisms (SNPs). Most of the pair-wise relative kinship coefficients (58.5%) were equal or close to 0, which suggests the lack of redundancy in the genomic composition in the majority of inbred lines. Genetic distance among most pairs of lines (98.3%) varied from 0.20 to 0.34 as compared with just 1.7% of the pairs of lines that differed by <0.20, which suggests greater genetic variation even among sister lines. The overall average of 17% heterogeneity was observed in the panel indicated the need for further inbreeding in the high heterogeneous genotypes. The mean nucleotide diversity and frequency of polymorphic sites observed in the panel were 0.28 and 0.02, respectively. The model-based population structure, principal component analysis, and phylogenetic analysis revealed three to six groups with no clear patterns of clustering by centers-wise breeding lines, types of corn, kernel characteristics, maturity, plant height, and ear placement. However, genotypes were grouped partially based on their source germplasm from where they derived.

Gut Microbial Composition Differs Extensively among Indian Native Chicken Breeds Originated in Different Geographical Locations and a Commercial Broiler Line, but Breed-Specific, as Well as Across-Breed Core Microbiomes, Are Found.

Gut microbiota plays an important role in the health and performance of the host. Characterizations of gut microbiota, core microbiomes, and microbial networks in different chicken breeds are expected to provide clues for pathogen exclusion, improving performance or feed efficiency. Here, we characterized the gut microbiota of "finishing" chickens (at the end of production life) of indigenous Indian Nicobari, Ghagus, and Aseel breeds, originating from the Nicobari island, coastal India, and the Indian mainland, respectively, as well as a global commercial broiler line, VenCobb 400, using 16S rDNA amplicon sequencing. We found that diversity, as well as richness of microbiota, was higher in indigenous breeds than in the broiler line. Beta diversity analysis indicated the highest overlap between Ghagus and Nicobari breeds and a very low overlap between the broiler line and all indigenous breeds. Linear discriminant analysis effect size (LEfSe) revealed 82 breed- or line-specific phylotype operational taxonomic unit (OTU) level biomarkers. We confirm the presence of breed specific and across-breed core microbiomes. Additionally, we show the existence of breed specific complex microbial networks in all groups. This study provides the first (and comprehensive) insight into the gut microbiota of three indigenous breeds and one commercial broiler line of chickens reared without antimicrobials, and underscores the need to study microbial diversity in other indigenous breeds.

Development of sub-tropically adapted diverse provitamin-A rich maize inbreds through marker-assisted pedigree selection, their characterization and utilization in hybrid breeding.

Malnutrition has emerged as one of the major health problems worldwide. Traditional yellow maize has low provitamin-A (proA) content and its genetic base in proA biofortification breeding program of subtropics is extremely narrow. To diversify the proA rich germplasm, 10 elite low proA inbreds were crossed with a proA rich donor (HP702-22) having mutant crtRB1 gene. The F2 populations derived from these crosses were genotyped using InDel marker specific to crtRB1. Severe marker segregation distortion was observed. Seventeen crtRB1 inbreds developed through marker-assisted pedigree breeding and seven inbreds generated using marker-assisted backcross breeding were characterized using 77 SSRs. Wide variation in gene diversity (0.08 to 0.79) and dissimilarity coefficient (0.28 to 0.84) was observed. The inbreds were grouped into three major clusters depicting the existing genetic diversity. The crtRB1-based inbreds possessed high ß-carotene (BC: 8.72µg/g), ß-cryptoxanthin (BCX: 4.58µg/g) and proA (11.01µg/g), while it was 2.35µg/g, 1.24µg/g and 2.97µg/g in checks, respectively. Based on their genetic relationships, 15 newly developed crtRB1-based inbreds were crossed with five testers (having crtRB1 gene) using line × tester mating design. 75 experimental hybrids with crtRB1 gene were evaluated over three locations. These experimental hybrids possessed higher BC (8.02µg/g), BCX (4.69µg/g), proA (10.37µg/g) compared to traditional hybrids used as check (BC: 2.36 µg/g, BCX: 1.53µg/g, proA: 3.13µg/g). Environment and genotypes × environment interaction had minor effects on proA content. Both additive and dominance gene action were significant for proA. The mean proportion of proA to total carotenoids (TC) was 44% among crtRB1-based hybrids, while 11% in traditional hybrids. BC was found to be positively correlated with BCX (r = 0.68) and proA (r = 0.98). However, no correlation was observed between proA and grain yield. Several hybrids with >10.0 t/ha grain yield with proA content >10.0 µg/g were identified. This is the first comprehensive study on development of diverse proA rich maize hybrids through marker-assisted pedigree breeding approach. The findings provides sustainable and cost-effective solution to alleviate vitamin-A deficiency.

Nutrient intake, digestibility, and blood metabolites of goats fed diets containing processed jatropha meal.

This study was conducted to record the ideal source and level of alkali treatment to treat jatropha meal (JM) and to determine the effect of inclusion of variously processed JM (pJM) on nutrient intake, digestibility, blood metabolites and hormonal status in goats. The JM was treated with 10 g/kg sodium chloride and 5 g/kg calcium hydroxide. The content of phorbol ester and hemagglutination (HA) activity of JM and pJM were assessed. A feeding trial for 90 days was conducted in short-haired multipurpose goats (n = 15; five per group). The experimental animals were offered oat (Avena sativa) straw ad libitum throughout the experimental period of 90 days. Each group was assigned to one of the three diets, viz. R1--soybean meal, R2--sodium chloride (10 g/kg dry matter, DM), and R3--calcium hydroxide (5 g/kg DM), with pJM substituting 250 g/kg DM of crude protein (CP) of control (R1). At the end of the feeding period, digestion trial of 7 days was conducted. Blood samples were collected at the end of the experimental period to assess the blood metabolites and hormonal status. The phorbol ester and HA activity were reduced considerably in pJM. The intake of DM, organic matter, CP, and nitrogen-free extract were comparable among all the groups. However, the intake of ether extract was significantly higher in pJM-fed groups. The hemoglobin, packed cell volume, serum urea, triiodothyronine and testosterone contents decreased in R2 and R3 as compared to R1. Concentration of glucose and activity of serum glutamic pyruvic transaminase and lactate dehydrogenase increased (P < 0.01) in goats fed pJM. It was concluded that phorbol ester content and HA activity markedly decreased by processing JM with sodium chloride and calcium hydroxide. However, they were not reduced to the levels of safe feeding, as reflected in unusual values of blood metabolites among the experimental animals.

Effect of supplementing different concentrations of organic trace minerals on performance, antioxidant activity, and bone mineralization in Vanaraja chickens developed for free range farming.

An experiment was conducted to determine the performance, antioxidation activity, and bone mineral content in Vanaraja chickens fed diet supplemented with organic trace minerals (oTM) at reduced levels. A total of 360 day-old chicks were selected and distributed randomly into 60 battery brooder pens. A maize-soybean meal-based control diet was supplemented with inorganic trace minerals (iTM), i.e., Mn, Zn, Fe, and Cu at 50, 45, 40, and 7.5 mg/kg, respectively, and varying concentration of oTM, i.e., Zn, Mn, Cu, Fe, I, Se, and Cr at 45, 50, 7.5, 40, 2, 0.30, and 0.25 mg/kg (diet II); 33.75, 37.50, 5.63, 30.0, 1.50, 0.23, and 0.19 mg/kg (diet III); 22.5, 25.0, 3.75, 20.0, 1.0, 0.15, and 0.13 mg/kg (diet IV); 18.0, 20.0, 3.0, 16.0, 0.80, 0.12, and 0.10 mg/kg (diet V); and 13.5, 15.0, 2.25, 12.0, 0.60, 0.09, and 0.08 mg/kg (diet VI), respectively. Each diet was allotted randomly to ten replicates and fed ad libitum from 1 to 42 days of age. The body weight at 14, 28, and 42 days was not affected by reducing the supplementation of oTM concentration in the diets. Similarly, feed intake at 14 days of age was not affected but reduced significantly (P < 0.05) in the group fed diet IV (50% oTM) compared to that in the other groups. The higher feed conversion ratio and increased concentration of Ca, P, and trace minerals in tibia were evident in the group fed oTM-supplemented diets compared to the diet containing iTM. Activities of glutathione peroxidase and ferric reducing ability in plasma did not differ in the groups fed on lower concentration of oTM compared to those fed on diet I (control diet). Therefore, it is concluded that the dietary supplementation of trace minerals can be reduced greatly when supplemented as organic form without affecting growth and antioxidant status in Vanaraja chickens.

Effect of supplementing organic selenium on performance, carcass traits, oxidative parameters and immune responses in commercial broiler chickens.

An experiment was conducted to determine the effect of supplementing various concentrations (0, 100, 200, 300, or 400 µg/kg diet) of organic Se on growth performance, carcass traits, oxidative stress, and immune responses in commercial broiler chickens reared in open-sided poultry house under tropical climatic conditions. Each diet was fed ad libitum to eight replicates consisting of six birds in each pen from 1 to 42 d of age. Body weight gain and feed efficiency, and relative weight of liver, abdominal fat and ready to cook yields were not affected (p>0.05) by organic Se supplementation to broiler diets. Lipid peroxidation in plasma decreased, while activities of glutathione peroxidase and glutathione reductase in plasma increased (p<0.01) linearly with Se concentration in diet. The ratios between heterophyls and lymphocytes and relative weight of lymphoid organs (bursa, spleen, and thymus), and antibody production to Newcastle disease vaccination were not affected (p>0.05) by Se supplementation to broiler diets. However, the cell-mediated immunity (lymphocyte proliferation ratio) increased (p<0.01) linearly with dietary Se concentration. The results of the present study indicate that the supplementation of Se did not influence body weight and feed efficiency. However, supplementation of Se increased antioxidant status and lymphocyte proliferation in broiler chickens.

Intake, digestibility and nitrogen balance in Mithun (Bos frontalis) offered urea-treated paddy straw based feed blocks.

The aim of the study was to determine the effect of feeding feed blocks containing varying proportion of urea-treated paddy straw (UTPS) on dry matter (DM) intake (DMI), nutrient utilization and N balance in Mithun. For the purpose, four adult male Mithun (279.5 ± 8.2 kg) were selected and offered four experimental rations viz. R(1) (Napier fodder + concentrate at 60:40), R(2) (UTPS + concentrate at 50:50), R(3) (UTPS + concentrate at 60:40) and R(4) (UTPS + concentrate at 70:30) in 4 × 4 Latin square design. The DMI % of body weight was 2.59, 2.96, 2.85 and 2.77 and the DMI g kg(-1) W(0.75) was 107, 123, 118 and 115 in Mithun fed R(1), R(2), R(3) and R(4), respectively. The mean DMI was (P < 0.01) higher in animals fed R(2) and R(3) than R(1) and R(4), whereas the water intake was (P < 0.01) higher in Mithun fed R(2), R(3) and R(4) than R(1). The digestibility of DM, organic matter, crude protein, crude fibre, neutral detergent fibre and cellulose were (P < 0.05) higher in animals fed R(2), R(3) and R(4) than R(1). A positive N balance was observed in all the experimental animals, with higher (P < 0.05) values among the animals offered R(2), R(3) and R(4) than R(1). The digestible crude protein and total digestible nutrient intakes were higher (P < 0.05) in Mithun fed R(2) and R(3) than R(1) and R(4). It is concluded that the UTPS can be incorporated up to 70% to formulate the complete feed/feed block and can be used for feeding of Mithun under complete confinement system.

Development and validation of a sensitive radioimmunoassay for progesterone estimation in unextracted mithun (Bos frontalis) plasma.

The objective of the study was to develop and validate a simple, reliable, and highly sensitive radioimmunoassay (RIA) for progesterone determination in mithun (Bos frontalis) plasma. The RIA was carried out in 20 microL unextracted mithun plasma. The progesterone standards ranging from 2 to 500pg/20 microL/tube were prepared in charcoal-treated hormone-free plasma. The sensitivity of the RIA procedure was 2 pg progesterone/20 microL/tube, which corresponds to 0.1 ng/mL; the 50 percent relative binding sensitivity was seen at 32pg/20 microL/tube. Plasma volumes for the RIA, viz. 10 and 20 microL, did not influence the shape of standard curve, even though a slight drop in the counts was seen with higher plasma volumes. For the biological validation of the assay, three cyclic, three in early pregnancy, and two in late pregnancy mithuns were used. Blood samples collected at weekly intervals for 42 days, from all animals, were assayed for plasma progesterone. The peak level of progesterone was registered at day 14 (day 21 of sampling) of the estrous cycle and the lowest at estrus; the progesterone concentrations increased and decreased gradually as sampling time advanced, in early and late pregnancy, respectively, which confirm the biological validation of the RIA. The RIA avoids the troublesome and laborious plasma extraction procedures. In conclusion, the RIA developed for progesterone determination in mithun blood plasma is sufficiently reliable, simple, and sensitive enough to estimate progesterone in all physiological variations in mithun.

Twenty-four-hour secretion patterns of luteinizing hormone in mithuns (Bos frontalis).

A 24 h secretion pattern of luteinizing hormone (LH) was not available in mithun (Bos frontalis), a semi-wild ruminant. To characterize the 24 h LH profiles, six female mithun calves (age 7.8 +/- 0.5 months and 102.5 +/- 5.6 kg; group I) and six female mithuns averaging 25.4 months of age and 240 kg (group II) were selected from the National Research Centre on Mithun farm and were maintained in semi-intensive systems. Blood samples collected from all the animals at 30 min intervals for 24h were assayed for plasma LH. Plasma progesterone was also estimated in twice-a-week samples collected for 6-week period preceding each 24h sampling to assess whether any animal had begun ovarian cyclicity. The body weights of all animals were also recorded weekly during the 6-week period. LH patterns consisted of frequent pulses of varying amplitude. Luteinizing hormone pulses occurred at an average frequency of 0.28/h ( approximately 7 pulses/24 h) and 0.15/h ( approximately 3.5 pulses/24 h) for mithuns of groups II and I, respectively, the rate did not differ markedly among mithuns within each group but was significantly different between the groups. Similarly, the magnitude of LH secretory pulses did not vary among mithuns within the group but was significantly higher in group II than in group I animals. In group II, the LH peaks averaged 1.59 and 1.00 ng/ml in mithun having the highest and lowest LH peaks, respectively and the corresponding values for group I mithuns were 0.66 and 0.51ng/ml. Mithun with higher peak LH levels also had higher mean LH concentrations (P<0.05). The mithuns of group II had significantly higher plasma progesterone concentration (0.89 +/- 0.02 ng/ml) than those recorded in group I mithuns (0.26 +/- 0.01 ng/ml). Plasma progesterone profiles suggested that no animal reached puberty. In conclusion, there was higher LH secretion with higher pulsatility and greater amplitude in group II mithuns than exhibited in mithuns of group I and the prepubertal mithuns of group II were in approaching puberty, which were also indicated by their plasma progesterone profiles, critical body weight and age required to attain puberty, in addition to higher pulsatility of LH secretion.

Development and validation of a simple, sensitive, second antibody format enzyme immunoassay (EIA) for LH determination in mithun (Bos frontalis) plasma.

The objective of this study was to develop and validate a simple and highly sensitive enzyme immunoassay (EIA) for LH determination in mithun plasma on microtitreplates using the biotin-streptavidin amplification system and the second antibody coating technique. Biotin was coupled to LH and used to bridge between streptavidin-peroxidase and immobilized antiserum in competitive assay. The EIA was carried out directly in 20 microL mithun plasma. The LH standards ranging from 6.25 pg/well/20 microL to 400 pg/well/20 microL were prepared in hormone free plasma collected from a mithun on day 3 post calving. The sensitivity of EIA procedure was 6.25 pg/well LH, which corresponds to 0.31 ng/mL plasma; the 50 percent relative binding sensitivity was seen at 100 pg/well/20 microL. Plasma volumes for the EIA viz. 10 and 20 microL did not influence the shape of standard curve even though a slight drop in the OD450 was seen with higher plasma volumes. A parallelism test was carried out to compare the endogenous mithun plasma LH with bovine LH standards. It showed good parallelism with the bovine standard curve. For the biological validation of the assay, 3 mithuns were used. These were administered 10 microg i.v., with a synthetic analogue of GnRH (Buserelin-Acetate, Intervet, India) and blood samples were collected at 15 min intervals using indwelling jugular catheter beginning 1 h prior to GnRH injection till 8 h post injection. In all animals, sharp increases in LH concentrations were recorded post GnRH administration, which confirms the biological validation of the EIA. In conclusion, the EIA developed for LH determination in mithun blood plasma is sufficiently reliable, economical, and sensitive enough to estimate LH in all physiological variations in mithun.