A Review on In-vitro Haploid Production in Orchids

The aim of writing this paper is to review production of haploids in orchids in vitro. Haploids possess half number of chromosomes and do not undergo fertilization. In vitro conditions provide necessary nutrients and conditions that are required for growth of haploid plantlets. The natural breeding cycle of orchids is very slow as well as unpredictable. To reduce this time, the technique of in vitro haploid production is used. In vitro conditions can decrease or shorten the time required for juvenile period in Orchids. Haploid plants are produced from in vitro haploid culture. This technique is useful to produce homozygous pure lines and to increase the yield of a plant.

Cellulase production and efficient saccharification of biomass by a new mutant Trichoderma afroharzianum MEA-12

Background Cellulase plays a key role in converting cellulosic biomass into fermentable sugar to produce chemicals and fuels, which is generally produced by filamentous fungi. However, most of the filamentous fungi obtained by natural breeding have low secretory capacity in cellulase production, which are far from meeting the requirements of industrial production. Random mutagenesis combined with adaptive laboratory evolution (ALE) strategy is an effective method to increase the production of fungal enzymes. Results This study obtained a mutant of Trichoderma afroharzianum by exposures to N-methyl-N’-nitro-N-nitrosoguanidine (MNNG), Ethyl Methanesulfonate (EMS), Atmospheric and Room Temperature Plasma (ARTP) and ALE with high sugar stress. The T. afroharzianum mutant MEA-12 produced 0.60, 5.47, 0.31 and 2.17 IU/mL FPase, CMCase, pNPCase and pNPGase, respectively. These levels were 4.33, 6.37, 4.92 and 4.15 times higher than those of the parental strain, respectively. Also, it was found that T. afroharzianum had the same carbon catabolite repression (CCR) effect as other Trichoderma in liquid submerged fermentation. In contrast, the mutant MEA-12 can tolerate the inhibition of glucose (up to 20 mM) without affecting enzyme production under inducing conditions. Interestingly, crude enzyme from MEA-12 showed high enzymatic hydrolysis efficiency against three different biomasses (cornstalk, bamboo and reed), when combined with cellulase from T. reesei Rut-C30. In addition, the factors that improved cellulase production by MEA-12 were clarified. Conclusions Overall, compound mutagenesis combined with ALE effectively increased the production of fungal cellulase. A super-producing mutant MEA-12 was obtained, and its cellulase could hydrolyze common biomasses efficiently, in combination with enzymes derived from model strain T. reesei, which provides a new choice for processing of bioresources in the future.

Comparative study utilizing different post-breeding treatment regimens in cyclic Arabian mares

Background and Aim: Post-breeding treatment is the most common practice in the reproductive management of mares. Oxytocin, uterine lavage, and intrauterine (I/U) antibiotic are usually used as prophylactic therapy. This study aimed to determine the most efficient prophylactic treatment regimen among six treatment protocols applied during natural breeding of cyclic Arabian mares. Materials and Methods: The current study was conducted on cyclic Arabian mares that were subdivided into three age categories (n=968): Category I (5-10 years, n=380), Category II (11-15 years, n=361), and Category III (≥16 years, n=227). Six prophylactic treatments were applied after 4 h of breeding. According to the treatment regimen, treated mares (n=483) were divided into six treatment groups: A (n=80), treated with I/U antibiotic (1 g gentamicin); B (n=81), I/U lavage (normal saline 500 mL); C (n=83), intramuscular (I/M) oxytocin (10 IU); D (n=82), I/U antibiotic and I/M oxytocin; E (n=78), I/U lavage and I/M oxytocin; and F (n=79), I/U lavage with I/U antibiotic and I/M oxytocin. Non-treated mares were classified as controls (n=485). Ultrasonography was performed to monitor pregnant mares 30 and 60 days after mating, and mares were followed until foaling. Results: Pregnancy and foaling results reveals that in age Category I, treatment with oxytocin alone or oxytocin with I/U lavage showed the highest pregnancy and foaling rates (p<0.01). In age Category II, the highest pregnancy and foaling rates were observed in lavage treatment (p<0.01), whereas, in age Category III, the good pregnancy and foaling rates were monitored in treatment with oxytocin and I/U lavage (p<0.01). Conclusion: Treatment with systemic I/M oxytocin is ideal in early age group mares (5-10 years of age). However, irrespective of the age categories, all mares exhibited high pregnancy and foaling rates after treatment with systemic I/M oxytocin and I/U lavage with normal saline (0.9%) 4 h post-breeding.

ASSESSING IN-SITU SEMI-NATURAL BUTTERFLY BREEDING APPROACH OF ORNITHOPTERA CROESUS (PAPILIONIDAE) ON BACAN ISLAND, MALUKU UTARA, INDONESIA

Indonesia is rich in endemic species of flora and fauna. One of them is Ornithoptera croesus butterfly, which is endemic to North Maluku. Habitat degradation and trade of this species have caused the populations to decline. To avoid the collection of butterfly specimens from nature and to preserve their habitat and population in nature, a semi-natural butterfly breeding practice at Bacan Island was initiated in 2013. This research was conducted to assess the breeding approach for O. croesus lydius using a qualitative descriptive method. The assessment was based on these variables: the specific ecology of the butterfly; the suitability of the breeding site and development model; the utilization of larval host plants and butterfly nectar plants; and the establishment of the birdwing population at the site. The observations and results are presented here. Based on the assessment, the in-situ semi-natural breeding approach is one of the solutions for sustainable use of this protected species.

A FIVE-DECADE SYSTEMATIC REVIEW OF RESEARCH PROGRESS ON PRODUCTION AND MANAGEMENT OF SMALL RUMINANTS IN BANGLADESH

Background: Small ruminant animals (SRA) have been associated with humans since their domestication are of world significance as socioeconomic animals providing products (meat, milk, wool, hair) that are in growing demand and sustainability of rural economics and many ecosystems with poverty elevation and food security in rural people in developing world including Bangladesh. People clearly want the benefit of SRA with the application of knowledge of science and technological research. Some research progress on production and management of SRA has been made during the past five decades in Bangladesh. However, such research information is often fragmented and not easily accessible, although these research findings are essential for further research and effective SRA development planning. Objective: This review aimed to improve the information based on various aspects of production and management of SRA by compiling and reviewing published research findings to identify constraints of their production for practical and sustainable solutions. Materials and Methods: A systematic literature of review of research articles on production and management of SRA published mainly in journals from 1968 to early 2021 of Bangladesh has been reviewed. A total of 332 inland research articles on these aspects supported with 19 foreign related articles have been reviewed and analyzed. Results: Bangladesh has approximately 26.435 million goats and 3.607 million sheep. Among goat population, 90% Black Bengal goat (BBG), 8.0% Jamunapari (JP) and 2.0% crossbred goats. Ten phenotypic coat color characteristics of BBG with high performance with solid black color goats documented. Sheep and goats are considered polyestrous species, and the average gestation length lasts for 150 days and give birth twice a year under good management and nutrition. Most of the smallholder farmers (80.5-95%) are raised their goats in semi-intensive system and 47% rural women are involved in goat rearing. Approximately 96% goat farmers and 60% sheep farmers used roadside grass and tree leaves and only 4.0% goat farmers used cultivated fodder whereas 40% sheep farmers used cultivated and road side grass to feed their sheep. Daily supplement of 100-300g concentrate to grazing SRA improved the growth and carcass gain, productive and reproductive performances. Castrated goats at eight weeks of age showed a much higher growth rate (76.33g/day) than un-castrated (62.72g/day) goats. The productive and reproductive performances of SRA have been reviewed and discussed. The BBG has excellent reproductive efficiency but its lower live weight and milk yield encouraged for crossbreeding with exotic bucks. The meat and milk yield traits of crossbred improved but reproductive efficiency decreased in comparison to BBG. Selection breeding of BBG not only increased live weight but also improved carcass value and preserves the indigenous genetic resources from extinction of unplanned breeding. Similarly, the crossbreds between native ewes  exotic rams resulted positive effect on wool quality and live weight but a negative effect on prolificacy and lambing interval. Almost all SRA farmers depended on natural breeding system with some research data on the uses of AI and more than 70% farmers are being faced breeding service shortages. Major advances in methods of semen collection and evaluation, evaluation of male fertility, cryopreservation of sperm and estrous cycle control have been reported in both the goat and sheep in Bangladesh. In addition, knowledge of ovulation control, timing of insemination, gamete biology has also been reported in inland literature. The average slaughtering age of BBG reported to be approximately 12 months whereas in native Bengal sheep has been suggested to be 6 to 9 with an average of 8 months to get maximum return. The major challenges for SRA production are breeds, poor management, inadequate feeds, breeding bucks and rams and high neonatal mortality (kids 25.2%) and lambs (12.4%) in Bangladesh. Conclusions: The SRA rearing as smallholder farmers managed mostly semi-intensive system and requires improved feeding, provision of veterinary medical services, financial assistance, improve natural breeding and/or AI services and extension services to encourage and enhance production of sheep and goat at rural levels in Bangladesh. An improvement in the performance of SRA would directly improve the diet and standard of living of rural smallholder farmers in Bangladesh. Keywords: Systematic review, Five decades, Small ruminants, Production, Management, Bangladesh

Pointing Out Some Issues Regarding Reproduction Management in Murciano-Granadina Goats

Two of the most important problems in high-yielding dairy goat farms are early and accurate pregnancy diagnosis and the appropriate dry off of lactating does before the next kidding. The hypothesis posits that it could be possible to identify pregnant does through maximum progesterone milk levels at any time during the pregnancy, and that there is an optimal time to apply a lactation inhibitor to help dry off lactating does. Therefore, 114 Murciano-Granadina breed goats were used, from which 74 goats were inseminated at week 20 of lactation and samples of milk from pregnant and non-pregnant goats were taken at two-week intervals. The average maximum progesterone milk levels were higher outside the natural breeding season (40° latitude) than in the breeding season (11.6 ± 1.13 vs. 8.6 ± 1.02 ng/mL), although the levels from pregnant and non-pregnant goats were similar (10.85 ± 1.3 vs. 9.74 ± 1.6 ng/mL), except in the case of pregnancy with four foetuses (12.5 ± 1.3 ng/mL). Milk yield at mating does not affect fertility until a value of at least 3250 mL/day. Pregnancy started to affect milk yield up to the +7th week and was 59.9% lower in the +10th week after mating, so the use of lactation inhibitors could be more effective from this latter week. In conclusion, the results show that it is not possible to detect gestation in goats reliably through the maximum concentration of progesterone in milk at any time during lactation, except in the case of goats gestating four foetuses, that the milk yield of goats at mating does not affect fertility until a value of at least 3250 mL/day, and that from the 10th week post-mating, the application of lactation inhibitors would be optimal.

Females of the four-eyed frog, Pleurodema thaul (Anura, Leptodactylidae), respond behaviourally to conspecific male scent

Among amphibians, conspecific chemical communication has been widely studied in Caudata. Adult anurans, by contrast, have received less attention. Recently, it was shown that chemical scents are also relevant for adult anuran intraspecific communication. In this context, we evaluate whether females of the four-eyed frog (Pleurodema thaul) respond to conspecific male scents. We carried out a double choice experiment in a Y-maze. Females were repeatedly presented with the scents of several males versus distilled water. To extract the scent from males, we acoustically stimulated males and then used the water from their aquaria for the experiments. Our data suggest that females are capable of responding behaviourally to male scents, since they spent longer periods in the zones with male scent, rather than in zones with water. We propose that under natural breeding conditions, females of P. thaul may use either their chemical sense or chemical cues to facilitate their encounters with males.

Investigating Plasma Metabolomic Profiles at the Time of Weaning, Based on Pregnancy Outcome, in Bos Taurus heifers

The objective of this study was to identify differences in the metabolomic profiles of heifers at the time of weaning based on their subsequent pregnancy outcomes. Angus/Simmental-cross heifers (n = 36) were weighed at weaning and 10ml of blood was collected. The blood plasma and white blood cells (WBCs) were isolated. Heifers were then assessed 30 days prior to artificial insemination (AI) for weight, pelvic area (PA), body condition scores (BCS) and reproductive tract scores (RTS). Reproductively mature heifers underwent an AI program with estrus detection and were then exposed to a bull two weeks after AI. Heifers were categorized into three groups: pregnant by AI, pregnant by natural breeding or open after 3 consecutive estrous cycles. 9 heifers pregnant by AI and 11 open heifers were chosen for metabolomic profiling. Of the 20 heifers selected, there was no difference in weight at weaning (355.6±21.9 kg AI, 364.2±33.3 kg Open, p &gt;0.05), BCS (5.7±0.4 AI, 5.6±0.4 Open, P &gt;0.05), RTS (4.1±0.6 AI, 4.0±0.7 Open, P &gt; 0.05) or PA (173.7±10.8cm2 AI, 175.6±9.1cm2 Open, P &gt; 0.05). Ten metabolites were found to be significantly up or down regulated in the heifers who remained open after three consecutive cycles, when compared to those pregnant by AI. Alanine, cystine, lysine, methionine, tyrosine, tryptophan and valine were down regulated while glycerol, fructose-6-phosphate and ribulose-5-phosphate were found to be up regulated in open heifers (P &lt; 0.05). RNA isolated from WBCs was used to compare the expression of five inflammatory cytokines in the open and pregnant by AI groups (TNFα, IL6, CXCL5, POSTN and MCP1). Inflammatory cytokines were increased in all heifers that remained open after three consecutive cycles (p&lt; 0.05). In summary, the quantity of specific metabolites present within the blood plasma are different at weaning between heifers with differing reproductive potentials. This could potentially be used to develop an assay to aid in selecting replacement females.

Genetic differentiation over a small spatial scale in the smooth newt (Lissotriton vulgaris)

Understanding the distribution of genetic variation is central for both population biology and conservation genetics. Genetic population structure can be primarily affected by the species’ dispersal ability, which is assumed to be limited in many amphibians. In this study, we estimated allelic differentiation metrics and FST indices to investigate genetic variation among natural breeding ponds of smooth newts (Lissotriton vulgaris) over a small spatial scale. Based on six microsatellite loci, we found a small, but significant allelic differentiation among clusters of natural breeding ponds (i.e. ‘local regions’), which result was in line with the calculation of corresponding hierarchical FST values. Analysis of molecular variance also indicated significant between-region variation in the study area. Pairwise estimations showed that only the furthermost regions differed from each other in both differentiation measures, but this difference was not attributable to geographic distances between ponds. Our results provide evidence that hierarchical genetic structure can be characteristic to breeding ponds of smooth newts on a small spatial scale in their natural breeding habitat, but dispersal distance may be less limited than previously thought in these philopatric caudates.

Phenology of Antler Casting and Occurance of Late-Breeding in Nebraska White-Tailed Deer

White-tailed deer (Odocoileus virginianus) in Nebraska primarily breed in November (stimulated by photoperiod), but this season extends into December and January for unfertilized females and healthy fawns reaching the appropriate breeding weight by winter. Variation in the typical mating period can be attributed to geographic region, skewed sex ratios, and fawns reaching sexual maturity during their first year of birth. During April of 2019, we observed late season breeding by a male white-tailed deer in the central Platte River valley and documented late antler casting for this same late-copulating male. An additional male was observed in April of 2020 still retaining antlers within the region. To understand the irregularity of these observations relative to the central Platte River valley, we documented observations of early and late antler casting utilizing long-term cast antler collection and camera trap data. We were able to establish a baseline for antler casting phenology within this region to denote a shift in average casting timeframe (more than one month later than previously recorded in this region) and document early and the latest known antler casting within the literature. Herein, we detail the variation in antler casting phenology for white-tailed deer in this region, and the latest known occurrence of natural breeding in the state. Our findings help us better understand the life history of the Nebraska white-tailed deer population and can assist state wildlife biologists and property managers as they assess seasonal harvest regulations and adaptively manage the changing resource.