Potential Breeding Research Articles

Identification and genetic analysis of major gene ST3GAL4 related to serum alkaline phosphatase in chicken

Alkaline phosphatase (ALP) is a marker of osteoblast maturation and an important indicator of bone metabolism. The activity of ALP can reflect the bone metabolism and growth traits of animals, so the polymorphism affecting ALP expression deserves further study. In this study, we identified an SNP site in ST3GAL4 found by genome-wide association studies (GWAS) in previous studies, 8 SNPs were also identified by DNA sequencing. Interestingly, there were 4 SNPs (rs475471G > A, rs475533C > T, rs475621A > G, rs475647C > A) completely linked by haplotype analysis. Therefore, we selected a tag SNP rs475471G > A to further analyze the ALP level of different genotypes in Hubbard leg disease population and an F2 chicken resource population produced by Anka and Gushi chickens and carried out population genetic analysis in 18 chicken breeds. Association analysis showed that this QTL within ST3GAL4 was highly correlated with ALP level. The mutant individuals with genotype AA had the highest ALP level, followed by GA and GG carriers. The mutant individual carriers of AA and GA genotype had higher values for body weight (BW), chest width (CW), body slanting length (BSL), pelvis width (PW) at 4-week, the semi-evisceration weight (SEW), evisceration weight (EW) and Leg weight (LW) than GG genotypes. The amplification and typing of 4852 DNA samples from 18 different breeds showed GG genotype mainly existed in egg-type chickens and dual-type chickens, while the AA genotype was mainly distributed in commercial broilers and F2 resource population. The individual carriers of the AA genotype had the highest ALP and showed better growth performance. Besides, tissue expression analysis used Cobb broiler showed significant differences between different genotypes in the spleen and duodenum. Taken together, this was the first time to determine 9 SNPs within ST3GAL4 related to ALP in chickens, 4 of them were complete linkage with each other, which provides useful information on the mutation of ST3GAL4 and could predict the serum ALP level of chicken early and as an effective potential molecular breeding marker for chickens.

Cell Membrane Features as Potential Breeding Targets to Improve Cold Germination Ability of Seeds.

Cold stress breeding that focuses on the improvement of chilling tolerance at the germination stage is constrained by the complexities of the trait which involves integrated cellular, biochemical, hormonal and molecular responses. Biological membrane serves as the first line of plant defense under stress. Membranes receive cold stress signals and transduce them into intracellular responses. Low temperature stress, in particular, primarily and effectively affects the structure, composition and properties of cell membranes, which ultimately disturbs cellular homeostasis. Under cold stress, maintenance of membrane integrity through the alteration of membrane lipid composition is of prime importance to cope with the stress. This review describes the critical role of cell membranes in cold stress responses as well as the physiological and biochemical manifestations of cold stress in plants. The potential of cell membrane properties as breeding targets in developing strategies to improve cold germination ability is discussed using cotton (Gossypium hirsutum L.) as a model.

The Characteristics Probabilities and Risks Prediction of Containers as Potential Breeding Sites for Aedes Aegypti Mosquitoes

Characterization of the Aedes aegypti larval container is an important factor influencing mosquito reproduction patterns and is an important aspect of integrated vector control. The study purpose to identify the characteristics and predict the risk of water reservoirs that have the potential as a breeding place for the Aedes aegypti mosquito. The study was using descriptive with a quantitative approach. It was conducted in the West Banjarmasin District which consisted of 9 villages (Kuin Selatan, Kuin Cerucuk, Belitung Selatan, Pelambuan, Belitung Utara, Telaga Biru, Telawang, Teluk Tiram, and Basirih). The population was all households in West Banjarmasin District with a total of 10.003 houses. A sample of 100 houses was taken using proportional random sampling. The results showed that 77.8% of households use tap water sources and there are 44,4% use dark colored containers. Most of the households have a water reservoir in the house (88.9%) and in Teluk Tiram Village about 66.70% of the households drain the water reservoir for more than 7 days. Concluded that the characteristics of most water reservoirs were tap water sources, dark in color, located inside the house, and cleaned more than 7 days with the highest risk value was 73.13%.

Knockout of a papain-like cysteine protease gene OCP enhances blast resistance in rice.

Papain-like cysteine proteases (PLCPs) play an important role in the immune response of plants. In Arabidopsis, several homologous genes are known to be involved in defending against pathogens. However, the effects of PLCPs on diseases that afflict rice are largely unknown. In this study, we show that a PLCP, an oryzain alpha chain precursor (OCP), the ortholog of the Arabidopsis protease RD21 (responsive to dehydration 21), participates in regulating resistance to blast disease with a shorter lesion length characterizing the knockout lines (ocp-ko), generated via CRISPR/Cas9 technology. OCP was expressed in all rice tissues and mainly located in the cytoplasm. We prove that OCP, featuring cysteine protease activity, interacts with OsRACK1A (receptor for activated C kinase 1) and OsSNAP32 (synaptosome-associated protein of 32 kD) physically in vitro and in vivo, and they co-locate in the rice cytoplasm but cannot form a ternary complex. Many genes related to plant immunity were enriched in the ocp-ko1 line whose expression levels changed significantly. The expression of jasmonic acid (JA) and ethylene (ET) biosynthesis and regulatory genes were up-regulated, while that of auxin efflux transporters was down-regulated in ocp-ko1. Therefore, OCP negatively regulates blast resistance in rice by interacting with OsRACK1A or OsSNAP32 and influencing the expression profiles of many resistance-related genes. Moreover, OCP might be the cornerstone of blast resistance by suppressing the activation of JA and ET signaling pathways as well as promoting auxin signaling pathways. Our research provides a comprehensive resource of PLCPs for rice plants in defense against pathogens that is also of potential breeding value.

Improving Nitrogen Use Efficiency in Rice for Sustainable Agriculture: Strategies and Future Perspectives.

Nitrogen (N) is an important nutrient for the growth and development of rice. The application of N fertilizer has become one of the inevitable ways to increase rice yield due to insufficient soil N content. However, in order to achieve stable and high yield, farmers usually increase N fertilizer input without hesitation, resulting in a series of problems such as environmental pollution, energy waste and low production efficiency. For sustainable agriculture, improving the nitrogen use efficiency (NUE) to decrease N fertilizer input is imperative. In the present review, we firstly demonstrate the role of N in mediating root architecture, photosynthesis, metabolic balance, and yield components in rice. Furthermore, we further summarize the current agronomic practices for enhancing rice NUE, including balanced fertilization, the use of nitrification inhibitors and slow-release N fertilizers, the split application of N fertilizer, root zone fertilization, and so on. Finally, we discuss the recent advances of N efficiency-related genes with potential breeding value. These genes will contribute to improving the N uptake, maintain the N metabolism balance, and enhance the NUE, thereby breeding new varieties against low N tolerance to improve the rice yield and quality. Moreover, N-efficient varieties also need combine with precise N fertilizer management and advanced cultivation techniques to realize the maximum exploitation of their biological potential.

CRISPR/Cas9-Targeted Knockout of Rice Susceptibility Genes OsDjA2 and OsERF104 Reveals Alternative Sources of Resistance to Pyricularia oryzae

Rice genes OsDjA2 and OsERF104, encoding a chaperone protein and an APETELA2/ ethylene-responsive factor, respectively, are strongly induced in a compatible interaction with blast fungus, and also have function in plant susceptibility validated through gene silencing. Here, we reported the CRISPR/Cas9 knockout of OsDjA2 and OsERF104 genes resulting in considerable improvement of blast resistance. A total of 15 OsDjA2 (62.5%) and 17 OsERF104 (70.8%) T0 transformed lines were identified from 24 regenerated plants for each target and used in downstream experiments. Phenotyping of homozygous T1 mutant lines revealed not only a significant decrease in the number of blast lesions but also a reduction in the percentage of diseased leaf area, compared with the infected control plants. Our results supported CRISPR/Cas9-mediated target mutation in rice susceptibility genes as a potential and alternative breeding strategy for building resistance to blast disease.

Sheep in the Vineyard: Suitability of Different Breeds and Potential Breeding Objectives.

Simple SummaryThere is a large number of sheep breeds worldwide. Today, many of them are endangered, because there are very little market demands for the majority of breeds. The integration of sheep in viticultural systems offers a promising option to put sheep and unique breed characteristics in value. However, it is unknown which characteristics and breeds are best suited for this purpose. The lack of information leads to problems in the implementation of this so-called new integrated crop-livestock system. In our research, we addressed this challenge. We studied 26 breeds and tested their suitability for integration into common vineyards of Central Europe. Two breeds fulfill the most important requirement. Southdown and Shropshire, for the latter especially, the sheep of the shorter legged Danish type seem to be unable to stand on two legs. Their muzzle heights stay within a tolerable range without harming the foliage area of vines. Therefore, adult animals of both breeds seem suitable to take over important viticultural tasks during the growing season. A third breed, the Ouessant sheep, is suitable with some limitations.Protecting a breed of sheep is simple when there is demand for its breed traits, but new market options are often hard to find. In general, grazing sheep are able to take over some viticultural work. Here, we address a new and promising integrated crop-livestock system that involves the integration of sheep in the vineyard during the growing season. Using sheep in a vineyard entails opportunities but also risks, such as the current lack of information, specifically in relation to breed traits. In our survey, we evaluated 26 breeds for their suitability for grazing as long as possible in Central European vineyards during the growing season. First, the breed traits required were identified. Then, 94 flock book breeders were interviewed about specific breed traits. The height of a sheep’s muzzle is particularly important for assessing the suitability of a breed, as it defines the potential impact on the foliage area during the growing season. To determine the height of the muzzle, 179 flock book animals were measured. We found that the most important breeding objective for a new breed of sheep is the inability to stand on two legs. Adult animals of the breed Shropshire, and among these especially the shorter-legged Danish type, and Southdown, show a widespread inability to stand on two legs. Ouessant sheep are able to do so, yet are suitable with some limitations. Due to their extraordinarily small size, their reach is limited, as is their grazing performance. Thus, three of the 26 breeds studied here seem suitable for use in the most widespread vine training systems of Central Europe during the growing season. Targeted breeding could further improve the suitability of sheep for viticulture. Our findings could help to protect breeds and breed traits.

Breeding records of the birds of south-east Sulawesi, Indonesia: a collation of observations encompassing nearly 20 years of research in Wallacea

The islands of the Wallacean biodiversity hotspot support diverse and highly endemic bird communities, yet remain ornithologically poorly studied. There is a particular paucity of data regarding breeding biology for the region's birds. Here, we help to address this research gap by collating observations of breeding evidence made during nearly 20 years of field work between 1999 and 2018 in south-east Sulawesi and its offshore islands. Data were collected between April and September, with a special focus on July and August, which is the dry season across the southern half of Sulawesi. In total, we summarise 1,064 observations of potential breeding in 66 species, of which 27 are Wallacean endemics (including the Critically Endangered Maleo Macrocephalon maleo), and 39 species of wider range (including the Endangered Milky Stork Mycteria cinerea). Records include species with little or no previously published information on their breeding biology, such as Pygmy Hanging Parrot Loriculus exilis, Sulawesi Pitta Erythropitta celebensis and an undescribed Zosterops species, provisionally referred to as Wangi-wangi White-eye.

The mitochondria-localized protein OsNDB2 negatively regulates grain size and weight in rice

Grain size is a determinant of rice grain yield. In plants, mitochondria supply energy for cellular metabolism via the mitochondrial electron transport chain. Here we report that OsNDB2, which encodes a putative rotenone-insensitive type II NAD(P)H dehydrogenase (ND), negatively regulates grain size and weight in rice. Six ND members representing three major types of rice were identified, and the predicted OsNDB2 protein was localized to mitochondria. Contents of OsNDB2 transcripts were higher in young panicles and leaf blades. OsNDB2 overexpression reduced grain length, grain width, and 1000-grain weight and moderately influenced plant height, while knockout of OsNDB2 increased grain size and 1000-grain weight. Allelic mutations of OsNDB2 were associated with diverse grain appearances. Cellular observations revealed that variations in grain size of transgenic lines were caused by change in cell expansion but not cell proliferation in spikelet hulls. Our study sheds light on OsNDB2 function and provides a new potential breeding approach for increasing rice grain size and weight.

Comparative study of the effects of different soluble salts on seed germination of wild marigold (Tagetes minuta L.)

Wild marigold (Tagetes minuta L.) is an important aromatic plant species with a high industrial value in its essential oil. It has spread worldwide due to its wide adaptations. However, meager information is available regarding its salt tolerance. Understanding its seed germination behavior is crucial for establishing the plant species in new saline habitats. The present work focuses on the effect of five soluble salts (CaCl2, KCl, MgCl2, Na2SO4 and NaCl) on germination velocity, salinity tolerance index and germination percentage of wild marigold. Five concentrations (0, 50, 100, 150 and 200 mM) of the salts were given in a two-factor factorial, completely randomized design (CRD). The experiment was performed in a programmed culture room at 25 ± 2 ℃ under 16 h of light (PAR: 40 µmol m−2 s−1) and 8 h dark. Findings suggest that seeds of wild marigold exhibit the highest germination percentage (GP) of more than 95% in non-saline (control) conditions. The trend of germination inhibition due to different soluble salts was in the order: Na2SO4 > KCl > NaCl > CaCl2 > MgCl2. There was a significant decrease in germination velocity (0–40.56) and salinity tolerance index (0–96.67) with the elevated levels of salinity (all applied soluble salts). The dose resulting in a 50% decrease in seed germination, i.e., IC50 (inhibition concentration value) of 79.9 mM of solute salts, was obtained for wild marigold. There are strong indications that wild marigold can tolerate moderate salinity levels and can be appraised as a salt-tolerant aromatic crop. The present study's findings will open future horizons of salt tolerance in wild marigold by identifying potential salt tolerant breeding lines.

The Solanum torvum Transcription Factor StoWRKY6 Mediates Resistance against Verticillium Wilt

WRKY is a transcription factor family that has attracted much attention in recent studies of plant disease resistance, but there are few reports in the study of eggplant resistance to Verticillium wilt. Here, we retrieved an up-regulated WRKY transcription factor, StoWRKY6, from the transcriptome sequencing data of Solanum torvum response to Verticillium dahliae infection. Phylogenetic analyses revealed the highest homology species of StoWRKY6 in the WRKY family is Solanum melongena. Based on the quantitative real-time PCR analysis, StoWRKY6 was highly expressed in the roots but barely expressed in the leaves. Transient expressions of StoWRKY6 in Nicotiana benthamiana showed a nuclear localization. A virus-mediated gene silencing experiment indicated that the silencing of StoWRKY6 reduced the resistance to Verticillium wilt in Solanum torvum. To further verify the immune response function, we introduced StoWRKY6 into Nicotiana benthamiana using transient transformation technology and found obvious spots under UV light. In summary, these results showed that StoWRKY6 played an important role in the resistance to Verticillium wilt of Solanum torvum, which may function mainly by inducing an immune response. Our study provided strong evidence for the mechanism of eggplant resistance to Verticillium wilt and laid a foundation for the potential molecular breeding of eggplant disease resistance.

Use of geographical information systems (GIS) in assessing ecological profile, fish community structure and production of a large reservoir of Himachal Pradesh.

The present study demonstrates the spatial analysis and mapping of fish and different measures of environmental parameters and fish diversity of Pong reservoir, Himachal Pradesh, using Kriging spatial interpolation methods for geographical information system mapping. Seasonal data on environmental parameters, potential fish habitat and fish diversity was collected from lentic (dam), lentic (reservoir), transitional and lotic zone of the reservoir.. Important environmental parameters like water temperature, dissolved oxygen, electrical conductivity, water depth and transparency showed variations across the different zones of the reservoir. The sediment of the reservoir was sandy clay loam in nature as per texture analysis. Fish species richness, Shannon index and evenness index showed a similarity of the lotic and lentic (reservoir) zones of the reservoir. Six potential fish breeding grounds were identified in the reservoir indicating high conservation significance. The analysis of data showed a declining trend in fish production from 456.9 tonnes during the decade 1976-1987 to 347.91 tonnes during 2009-2020. The factors like anthropogenic climate change, predation of a stocked fish juvenile by water birds, undersized fish stocking and unscientific management are the probable reasons for the decreasing fish production. The spatial variation pattern of the water spread area, environmental parameters, fish catch and potential fish breeding grounds depicted in the GIS platform can be used as an important information base by the policy makers for fisheries management. The stocking of large size fish as a stocking material and adequate protection of the potential fish breeding grounds are the key advisories for the sustainable enhancement of fisheries as well as conservation.

Climate change is predicted to reduce sympatry among North American wood-warblers

Abstract Anthropogenic climate change will dramatically alter species distributions. The rate and magnitude of range shifts, however, will differ among taxa, resulting in altered patterns of co-occurrence and interspecific interactions. We examined potential climate-mediated breeding range shifts among North American wood-warblers (Parulidae), a speciose avian family likely to be especially impacted by such changes due to high levels of interspecific competition and hybridization. We used publicly available species distribution model (SDM) range outputs to compare current ranges and patterns of sympatry among warbler species to future ranges and sympatry under 1.5°C, 2.0°C, and 3.0°C of average global warming. Range overlap among species and number of sympatric species are expected to decrease significantly in future warming scenarios, and unequal range shifts will alter the composition of warbler communities. On average, climate change will result in net decreases in the local species diversity; each warbler species is predicted to gain sympatry with approximately 1 new species and lose sympatry with approximately 2 species. Climate-mediated changes are predicted to differ among warblers in different regions of North America, with greatest impacts on eastern and boreal forest species. Our findings suggest that climate change will alter the diversity of wood-warbler communities during this century. Targeted monitoring of these changing interspecific relationships, especially for antagonistic interactions or hybridization between newly sympatric species, will be crucial for prioritizing particular species and regions in future conservation or management efforts.

Selecting putative drought-tolerance markers in two contrasting soybeans

Identifying high-yield genotypes under low water availability is essential for soybean climate-smart breeding. However, a major bottleneck lies in phenotyping, particularly in selecting cost-efficient markers associated with stress tolerance and yield stabilization. Here, we conducted in-depth phenotyping experiments in two soybean genotypes with contrasting drought tolerance, MUNASQA (tolerant) and TJ2049 (susceptible), to better understand soybean stress physiology and identify/statistically validate drought-tolerance and yield-stabilization traits as potential breeding markers. Firstly, at the critical reproductive stage (R5), the molecular differences between the genotype’s responses to mild water deficit were explored through massive analysis of cDNA ends (MACE)-transcriptomic and gene ontology. MUNASQA transcriptional profile, compared to TJ2049, revealed significant differences when responding to drought. Next, both genotypes were phenotyped under mild water deficit, imposed in vegetative (V3) and R5 stages, by evaluating 22 stress-response, growth, and water-use markers, which were subsequently correlated between phenological stages and with yield. Several markers showed high consistency, independent of the phenological stage, demonstrating the effectiveness of the phenotyping methodology and its possible use for early selection. Finally, these markers were classified and selected according to their cost-feasibility, statistical weight, and correlation with yield. Here, pubescence, stomatal density, and canopy temperature depression emerged as promising breeding markers for the early selection of drought-tolerant soybeans.

Recent Advances in DNA Methylation and Their Potential Breeding Applications in Plants

Traditional plant breeding encompasses repetitive crossing and selection based on morphological traits, while phenotypic selection has been complemented by molecular methods in recent decades. Genome editing with techniques like the CRISPR-Cas9 system is still a novel approach that is being used to make direct modifications to nucleotide sequences of crops. In addition to these genetic alterations, an improved understanding of epigenetic variations such as DNA methylation on the phenotype of plants has led to increased opportunities to accelerate crop improvement. DNA methylation is the most widely studied epigenetic mark in plants and other eukaryotes. These epigenetic marks are highly conserved and involved in altering the activities and functions of developmental signals by catalyzing changes in the chromatin structure through methylation and demethylation. Cytosine methylation (5mC) is the most prevalent modification found in DNA. However, recent identification of N6-methyladenosine (6mA) in plants starts to reveal their critical role in plant development. Epigenetic modifications are actively involved in creating the phenotype by controlling essential biological mechanisms. Epigenetic modifications could be heritable and metastable causing variation in epigenetic status between or within species. However, both genetic and heritable epigenetic variation has the potential to drive natural variation. Hence, epigenome editing might help overcome some of the shortcomings of genome editing (such as gene knockout), which can have significant off-target effects and only enables the loss of a gene’s function. In this review, we have discussed the mechanism underlying DNA methylation and demethylation in plants. Methyltransferases and demethylases are involved in catalyzing specific types of modification. We also discuss the potential role of DNA modifications in crop improvement for meeting the requirements of sustainable and green agriculture.

Projected Climate and Hydroregime Variability Constrain Ephemeral Wetland-Dependent Amphibian Populations in Simulations of Southern Toads.

Amphibian populations are threatened globally by stressors, including diminishing availability of suitable wetland breeding sites, altered hydroregimes driven by changing weather patterns, and exposure to contaminants. Ecological risk assessment should encompass spatial and temporal scales that capture influential ecological processes and demographic responses. Following the PopGUIDE framework of population model development for risk assessment, we used matrix population models, in conjunction with existing hydroregime predictions, under a climate change scenario to evaluate the effects of environmental stochasticity and aquatic pesticide exposure on amphibians that are dependent on ephemeral wetlands. Using southern toads (Anaxyrus terrestris) as an example, we simulated population dynamics with breeding success dependent on hydroregime suitability. Years were defined as optimal, marginal, or insufficient for successful toad recruitment, based on the duration of their potential breeding season and rate of larval development to metamorphosis. We simulated both probabilistic and chronologically specific population projections, including variable annual fecundity, based on hydroregime suitability and reduced larval survival from carbaryl exposure. In our simulations, populations were more negatively impacted by prolonged drought, and consequently multiple sequential years of reproductive failure, than by aquatic pesticide exposure. These results highlight the necessity of reliable climate projections to accurately represent the effects of altered hydroregimes on amphibian populations. Risk assessment approaches could be improved with flexible modifications that allow inclusion of various extrinsic stressors and identification of demographic and ecological vulnerabilities when precise data are lacking.

Record of a juvenile Omura's whale live stranded in Hainan Island: Evidence of a potential breeding ground in the South China Sea?

On January 12, 2011, a very young juvenile Bryde's-like baleen whale was found stranded alive on the east coast of Hainan Island, in the South China Sea. Unfortunately, it died the next morning. This baleen whale showed some unique external morphology and skull features of the Omura's whale (Balaenoptera omurai). Subsequently, it was genetically identified as an Omura's whale using two different mitochondrial DNA markers. This study documents the first confirmed live stranding of a juvenile Omura's whale on Hainan Island, China. Our findings expand the known distribution range of this species in Chinese waters and may indicate that waters around Hainan Island in the South China Sea could be a potential breeding ground for the Omura's whale.

Predicting the breeding hotspots of the southern right whale, Eubalaena australis (Cetartiodactyla: Mysticeti), along the southern Brazilian coastline

To predict the potential breeding hotspots of southern right whales in southern Brazil, we used a Maxent model with a “presence-only” dataset. The dataset was obtained from a systematic aerial survey and public records of the Programa de Monitoramento de Cetáceos da Bacia de Santos and Sistema de Apoio ao Monitoramento de Mamíferos Marinhos. Because of spatial autocorrelation, 528 records out of 3028 were used in the modelling process. The explanatory variables used in the models were coastal distance, linearity, slope and substrate type. The models were created separately for unaccompanied adults (Ad) and mother-calf pairs (Fe). Both models showed good accuracy according to their area under the curve values (Ad=0.974; Fe=0.958). Coastal distance was the most relevant explanatory variable for the unaccompanied adult model (54%), whereas coastal linearity was more relevant for the mother-calf pair model (82%). The estimated area for mother-calf pairs was more restricted to coastal areas than that for unaccompanied adults, possibly owing to the high number of shelter areas for calves near the coast. This is the first study to predict the potential breeding hotspots of southern right whales in Brazil and its results will allow for a more directed management of these whales and provide further research opportunities.

Prevalence of alcohol-tolerant and antibiotic-resistant bacterial pathogens on public hand sanitizer dispensers

BackgroundSince the advent of the COVID-19 pandemic, alcohol-based hand sanitizer dispensers (HSDs) have been installed in most public and clinical settings for hygiene purposes and convenient application.AimTo determine whether sanitizer-tolerant bacterial pathogens can colonize HSDs, spreading diseases and antibiotic resistance.MethodsSampling was conducted from operational automatic HSDs, specifically the dispensing nozzle in direct contact with sanitizer. Culture-dependent cultivation of bacteria and MALDI-TOF were employed to assess microbiological contamination. Bacterial isolates were selected for rapid killing and biofilm eradication assays with alcohol treatment. Antibiotic minimum inhibitory concentration assays were performed according to the Clinical and Laboratory Standards Institute guidelines. Virulence potential of bacterial isolates was evaluated in the Caenorhadbitis elegans infection model.FindingsNearly 50% of HSDs from 52 locations, including clinical settings, food industry, and public spaces, contain microbial contamination at 103–106 bacteria/mL. Bacterial identification revealed Bacillus cereus as the most frequent pathogen (29%), while Enterobacter cloacae was the only Gram-negative bacterial pathogen (2%). Selecting B. cereus and E. cloacae isolates for further evaluation, these isolates and associated biofilms were found to be tolerant to alcohol with survival up to 70%. They possessed resistance to various antibiotic classes, with higher virulence than laboratory strains in the C. elegans infection model.ConclusionHSDs serve as potential breeding grounds for dissemination of pathogens and antibiotic resistance across unaware users. Proper HSD maintenance will ensure protection of public health and sustainable use of sanitizing alcohols, to prevent emergence of alcohol-resistant pathogens.

Genetics of Base Coat Colour Variations and Coat Colour-Patterns of the South African Nguni Cattle Investigated Using High-Density SNP Genotypes.

Nguni cattle are a Sanga type breed with mixed B. taurus and B. indicus ancestry and proven resistance to ticks, diseases and other harsh conditions of the African geographical landscape. The multi-coloured Nguni coats have found a niche market in the leather industry leading to breeding objectives towards the promotion of such diversity. However, there is limited studies on the genomic architecture underlying the coat colour and patterns hampering any potential breeding and improvement of such trait. This study investigated the genetics of base coat colour, colour-sidedness and the white forehead stripe in Nguni cattle using coat colour phenotyped Nguni cattle and Illumina Bovine HD (770K) genotypes. Base coat colour phenotypes were categorised into eumelanin (n = 45) and pheomelanin (n = 19). Animals were categorised into either colour-sided (n = 46) or non-colour-sided (n = 94) and similarly into presence (n = 15) or absence (n = 67) of white forehead stripe. Genome-wide association tests were conducted using 622,103 quality controlled SNPs and the Efficient Mixed Model Association eXpedited method (EMMAX) implemented in Golden Helix SNP Variation Suite. The genome-wide association studies for base coat colour (eumelanin vs. pheomelanin) resulted into four indicative SNPs on BTA18 and a well-known gene, MC1R, was observed within 1 MB from the indicative SNPs (p < 0.00001) and found to play a role in the melanogenesis (core pathway for melanin production) and the MAPK signalling pathway. GWAS for colour-sidedness resulted in four indicative SNPs, none of which were in close proximity to the KIT candidate gene known for colour-sidedness. GWAS for the white forehead stripe resulted in 17 indicative SNPs on BTA6. Four genes MAPK10, EFNA5, PPP2R3C and PAK1 were found to be associated with the white forehead stripe and were part of the MAPK, adrenergic and Wnt signalling pathways that are synergistically associated with the synthesis of melanin. Overall, our results prove prior knowledge of the role of MC1R in base coat colours in cattle and suggested a different genetic mechanism for forehead stripe phenotypes in Nguni cattle.