Effects of long-term contrasting lime and phosphorus applications on barley grain yield,root growth and abundance of mycorrhiza

Lime and phosphorus (P) applications are common agricultural management practices. Our aim was to quantify the effects of long-term application practices on root growth and abundance of arbuscular mycorrhizal fungi (AMF) under field conditions. We assessed the effects of lime and P fertilizer applications on barley yield, root growth and AMF abundance in 2016. Treatments were no, low, medium and high liming rate corresponding to application of 0, 4, 8 and 12 Mg lime ha⁻¹ every 5–9 years since 1942 combined with no or yearly application of 15.6 kg P ha⁻¹ since 1944. At harvest, grain yield, root intensity (core-break) and AMF abundance at different soil depths were estimated. Root development was monitored during early growth with minirhizotrons in treatments receiving low, medium and high liming rates and P fertilization. A quadratic model relating grain yield to liming rate estimated yields to peak at 6.4 Mg lime ha⁻¹ with yields of 4.2 and 3.2 Mg grain ha⁻¹ with and without P fertilization, respectively. Low and medium liming rates resulted in greater AMF abundance, especially in the no P treatments. During early growth in P-fertilized treatments, 77% and 65% more roots developed in the soil profile when treated with medium and high liming rate, respectively, compared to low liming rate. We conclude that long-term application of lime in soils receiving yearly P fertilization improved conditions for root growth in soil layers below 30 cm, but at the high liming rate, this did not translate into higher yield.     

Development and characterization of five novel cell lines from snubnose pompano,Trachinotus blochii (Lacepede, 1801), and their application in gene expression and virological studies

Five novel permanent cell lines have been established from gill, heart, kidney, eye and fin of snubnose pompano, Trachinotus blochii. They were designated as snubnose pompano gill (SPG), snubnose pompano heart (SPH), snubnose pompano kidney (SPK), snubnose pompano eye (SPE) and snubnose pompano fin (SPF), respectively. All these cell lines were characterized and cryopreserved successfully at different passage levels. Cell lines were passaged every alternate day; SPG, SPH, SPK, SPE and SPF cell lines attained passage levels of 68, 74, 82, 79 and 106, respectively, since the initiation of their development in 2019. The cell lines grew well in Leibovitz's 15 medium containing 15% foetal bovine serum at 28°C. Immunophenotyping of the cell lines revealed the presence of fibronectin and pancytokeratin. No mycoplasma contamination was found. The transfection study revealed the gene expression efficiency of these cell lines by expressing the green fluorescent protein (GFP). The authentication on origin of cell lines from T. blochii was confirmed by amplification of species-specific mitochondrial cytochrome oxidase I gene. The results showed the susceptibility of these cell lines to fish nodavirus (FNV) and tilapia lake virus (TiLV) and resistance to cyprinid herpesvirus 2 (CyHV-2). The FNV infection in the cell lines was confirmed by RT-PCR, Western blot, ELISA and immunocytochemistry, while TiLV infection was confirmed by RT-PCR assay. These results revealed that these cell lines are suitable for virological and foreign gene expression studies.     

Effects of pyrethroid insecticide exposure on haematological and haemostatic profiles in rats

High malaria burden has led to the increase use of insecticides in the tropics and subtropics. This study thus aimed at assessing the haematological effects and associated haemostatic alteration of pyrethroid insecticide exposure using experimental animal model. Rats of comparable ages and weights were randomized into four groups (A-D). Rats in groups B, C and D were exposed to pyrethroid insecticide by inhalation for 1, 2 and 3 min daily respectively for three weeks. Rats in group A (control) were not exposed. Haematological and haemostatic variables were comparable in all groups (< 0.05). Results from the study show that minimal exposure to pyrethroids is safe.     

Effects of reaggregated granulosa cells and oocytes derived from early antral follicles on the properties of oocytes grown in vitro

In this study, we examined the effects of reconstructed oocyte–granulosa cell complexes (OGCs) on the development of porcine oocytes derived from early antral follicles (EAFs; 0.5–0.7 mm in diameter). When denuded oocytes were cocultured with granulosa cells derived from other EAFs, the oocytes and granulosa cells aggregated to form OGCs after 2 days of culture. After 14 days of culture, we compared cell number, oocyte diameter, and oocyte chromatin configuration in unmanipulated (natural) OGCs, reconstructed OGCs, and OGCs collected from antral follicles (AFs, 3.0–6.0 mm in diameter). The diameters of oocytes from reconstructed OGCs grown in vitro were not different from those of oocytes from natural OGCs, although they were significantly smaller than those of oocytes from antral follicle (AF) OGCs. Oocyte chromatin configuration did not differ among the 3 OGC groups, but the oocyte nuclear maturation rate was lower in the reconstructed OGCs and higher in the AF OGCs. However, when the in vitro culture period for the reconstructed OGCs was extended by 2 days, the nuclear maturation rate of oocytes from reconstructed OGCs was similar to that of oocytes from natural OGCs. In addition, blastocysts were successfully obtained from oocytes from reconstructed OGCs. In conclusion, we established an innovative culture method that allows oocytes and granulosa cells from EAFs to reaggregate as reconstructed OGCs, which yield oocytes with the ability to develop to the blastocyst stage.     

Population distribution and threats to sustainable management of selected non-timber forest products in tropical lowland rainforests of south western Nigeria

Uncontrolled harvesting of non-timber forest products (NTFPs) poses a serious risk of extermination to several of these species in Nigeria. Yet, there is a paucity of information on the distribution, population status and sustainable management of NTFPs in most of the tropical lowland rainforests. We, therefore, assessed the population, distribution and threats to sustainable management of NTFPs within the tropical lowland rainforests of Omo and Shasha Forest Reserves, south western Nigeria. Data were obtained through inventory surveys on five top priority species including: bush mango (Irvingia gabonensis (Aubry-Lecomte ex O’Rorke) Baill), African walnut (Tetracarpidium conophorum (Mull. Arg.) Hutch. & Dalziel syn. Plukenetia conophora), chew-stick (Massularia acuminata (G. Don) Bullock), fever bark (Annickia chlorantha Setten & P.J.Maas syn. Enantia chloranta) and bush pepper (Piper guineense Schumach. & Thonn.). Purposive and stratified random sampling techniques were used for the inventory. Each forest reserve was stratified into three, viz: less disturbed natural forest (for areas that have been rested for at least ten years), recently disturbed natural forest (for areas that have suffered one form of human perturba- tion or the other in the last five years), and plantation forest (for areas carrying forest plantation). Data were collected from eighteen 10m×500m belt transects located in the above strata. The species were gener- ally fewer in both plantation and recently disturbed natural forest than the less disturbed natural forest, suggesting that forest disturbances (habitat modification) for other uses may have an effect on the occurrence and densities of the NTFPs. Exceptions to this trend were found for P. guineense and T. conophorum, which were fairly common in both planta-tion and recently disturbed natural forest. Among three tree NTFP species (i.e. I. gabonensis, M. acuminata and A. chlorantha), only I. gabonensis showed a significant difference in overall DBH size classes for both reserves (t=-2.404; df =21; p=0.026). Three tree NTFP species in both reserves further showed differences from the regular patterns of distribution of trees. The fairly regular reverse J-shaped size class distribution observed for M. acuminata in the study sites, however, suggests a recuperating population. In general, destructive harvesting of species, logging operations, low population size, narrow distribution ranges and habitat degradation are the major threats to the population of NTFPs in the study area. The implications of our findings for sustainable management of NTFPs in the study area are discussed and recommendations are made for a feasible approach towards enhancing the status of the species.     

Sexing of Dog Sperm by Fluorescence In Situ Hybridization

Effective preselection of sex has been accomplished in several species of livestock and also in humans using the flow cytometric sperm sorting method. A guaranteed high sorting accuracy is a key prerequisite for the widespread use of sperm sexing. The standard validation method is flow cytometric remeasurement of the DNA content of the sexed sperm. Since this method relies on the same instrument that produced the original sperm separation, it is not truly independent. Therefore, to be able to specifically produce either male or female offspring in the dog, we developed a method of direct visualization of sex chromosomes in a single sperm using fluorescence in situ hybridization (FISH) as a validation method. Denaturation of canine spermatozoa by immersion in 1 M NaOH for 4 min yielded consistent hybridization results with over 97% hybridization efficiency and a good preservation of sperm morphology. There was no significant difference between the theoretical ratio (50:50) and the observed ratio of X- and Y-chromosome-bearing spermatozoa in any of the three dogs. In addition, the mean purities of flow-sorted sex chromosomes in spermatozoa of the three dogs were 90.8% for the X chromosome fraction and 89.6% for the Y chromosome fraction. This sorting was evaluated by using the dual color FISH protocol. Therefore, our results demonstrated that the FISH protocol worked reliably for both unsorted and sexed sperm samples.     

Pathological and Bacteriological Studies of Hydrosalpinx in Buffaloes

The objectives of the present study were to investigate the bacteria accompanying hydrosalpinx of the buffalo cow and investigate the correlation between bacterial infection of the uterus and hydrosalpinx. Buffalo cows’ reproductive tracts were collected from Mosul abattoir. A total 385 uterine samples were examined of which 25 were having hydrosalpinx. Swabs for bacteriology, fluid for cytology and biopsies for histopathology were collected from the hydrosalpinx and the uterus from each samples included in this study. Results of this study indicated high prevalence of hydrosalpinx (6.5%) including unilateral (n = 19; 76%) and bilateral (n = 6; 24%) hydrosalpinx. Although 16 samples (64%) of the hydrosalpinx samples had no bacterial growth, the most prevalent bacteria recovered from hydrosalpinx were Corynebacterium hemolyticum and Actinomyces bovis, 42.8% and 28.6%, respectively. The most prevalent bacteria in the uterus were Archanobacterium pyogenes (18.5%), Staphylococcus aureus (14.8%), and Listeria monocytogenes (11.0%). Higher rates of leukocytes infiltration (p < 0.01) were observed in the uterine discharge than hydrosalpinx. A significant (p < 0.01) increase in lymphocytes was found in uterine discharge. Microscopic examination of the hydrosalpinx showed mucosal atrophy and dilatation of oviductal lumen without any signs of inflammation. It could be concluded that there is no correlation between bacteria isolated from uterus and hydrosalpinx. No association was found between bacteriological cultures and hydrosalpinx. Inflammation of the uterine tissue could be extended to utero‐tubal junction producing local inflammation resulting in fibrosis and tubal obstruction. The obstruction in the lumen of the oviducts resulted in accumulation of fluid.