Expression and localization of angiopoietin family in corpus luteum during different stages of oestrous cycle and modulatory role of angiopoietins on steroidogenesis,angiogenesis and survivability of cultured buffalo luteal cells

The objective of this study was to document the expression and localization of angiopoietin (ANGPT) family members comprising of angiopoietin (ANGPT1 and ANGPT2), and their receptors (Tie1 and Tie2) in buffalo corpus luteum (CL) obtained from different stages of the oestrous cycle, and the modulatory role of ANGPT1 and ANGPT2 alone or in combinations on progesterone (P₄) secretion and mRNA expression of phosphotidylinositide‐3kinase‐protein kinase B (PI3K‐AKT), phosphoinositide‐dependent kinase (PDK), protein kinase B (AKT), Bcl2 associated death promoter (BAD), caspase 3 and von willebrand factor (vWF) in luteal cells obtained from midluteal phase (MLP) of oestrous cycle in buffalo. Real‐time RT‐PCR (qPCR), Western blot and immunohistochemistry were applied to investigate mRNA expression, protein expression and localization of examined factors whereas, the P₄ secretion was assessed by RIA. The mRNA and protein expression of ANGPT1 and Tie2 was maximum (p < .05) in mid luteal phase (MLP) of oestrous cycle. The ANGPT2 mRNA and protein expression was maximum (p < .05) in early luteal phase, decreased in MLP and again increased in late luteal phase of oestrous cycle. ANGPT family members were localized in luteal cells and endothelial cells with a stage specific immunoreactivity. P₄ secretion was highest (p < .05) with 100 ng/ml at 72 hr when luteal cells were treated with either protein alone. The mRNA expression of PDK, AKT and vWF was highest (p < .05) and BAD along with caspase 3 were lowest (p < .05) at 100 ng/ml at 72 hr of incubation period, when cultured luteal cells were treated with either protein alone or in combination. To conclude, our study explores the steroidogenic potential of angiopoietins to promote P₄ secretion, luteal cell survival and angiogenesis through an autocrine and paracrine actions in buffalo CL.     

Factors influencing the quality of half‐pearls (mabé) produced by the winged pearl oyster,Pteria penguin (Röding, 1758)

The winged pearl oyster, Pteria penguin, is cultured primarily to produce half‐pearls (mabé). The mabé quality is influenced by culture techniques, but there is limited information in this field. P. penguin with mean (±SE) dorso‐ventral height of 250 ± 6.5 mm were used to investigate the influence of culture period and nucleus position on mabé quality. Oysters were relaxed using 1‐propylene phenoxetol, and five nuclei were glued at different positions to the inner surfaces of the oyster shells; three on the more concave left valve and two on the right valve. Nucleated oysters were then cultured for 10 months under commercial pearl farming conditions at Savusavu in Fiji. Nacre deposited at the base and top of the nuclei was measured monthly, from the 6th to the 10th months of culture and the different qualities of mabé produced at different positions were scrutinized. Nacre thicknesses at the base and top of the resulting mabé were significantly different at different months (P < 0.05) and the rate of nacre deposition was highest during the warmer months. The different positions of nuclei on the valve greatly affected the quality of mabé formed. After a 10‐month culture period, around 1 mm of nacre covered the nuclei although the best quality mabé were obtained after 9 months.     

Morphologic and biochemical characterization of hyperextension of the metacarpophalangeal and metatarsophalangeal joints in llamas

OBJECTIVE: To determine the morphologic and biochemical characteristics of hyperextension of the metacarpophalangeal and metatarsophalangeal joints in llamas. ANIMALS: 12 adult llamas (6 with bilateral hyperextension of the metacarpophalangeal or metatarsophalangeal joints and 6 age- and sex-matched control llamas). PROCEDURES: Llamas were evaluated by use of lameness examination, ultrasonography, and radiography. A CBC, serum biochemical analysis, and determination of concentrations of trace minerals in serum and liver samples were performed. Llamas were euthanized, and samples of the superficial digital flexor tendon, deep digital flexor tendon, and suspensory ligament were obtained from 4 areas and snap-frozen in liquid nitrogen or suspended in neutral-buffered 10% formalin. Immunohistochemical evaluation of collagen types I and III and assays for measurement of lysyl oxidase activity were performed. RESULTS: 2 affected llamas had a visible gait deficit associated with metacarpophalangeal joint hyperextension. Radiographic evidence of osteoarthritis was detected in 1 severely affected llama, and ultrasonographic changes of soft tissue mineralization and suspensory desmitis were observed in 2 llamas. Liver concentrations of copper were lower and serum concentrations of zinc higher in affected llamas, compared with values in control llamas. Lysyl oxidase activity and collagen distribution did not differ significantly between groups. CONCLUSIONS AND CLINICAL RELEVANCE: Hyperextension of the metacarpophalangeal or metatarsophalangeal joints in llamas does not appear to be the result of injury or degeneration of the suspensory ligament or flexor tendons. Lower copper concentrations coupled with higher zinc concentrations in affected llamas may be indicative of secondary copper deficiency.     

Ace and ace-like genes of invasive redlegged earth mite: copy number variation,target-site mutations,and their associations with organophosphate insensitivity

Background

Invasive Australian populations of redlegged earth mite, Halotydeus destructor (Tucker), are evolving increasing organophosphate resistance. In addition to the canonical ace gene, the target gene of organophosphates, the H. destructor genome contains many radiated ace-like genes that vary in copy number and amino acid sequence. In this work, we characterise copy number and target-site mutation variation at the canonical ace and ace-like genes and test for potential associations with organophosphate insensitivity. This was achieved through comparisons of whole-genome pool-seq data from alive and dead mites following organophosphate exposure.

Results

A combination of increased copy number and target-site mutations at the canonical ace was associated with organophosphate insensitivity in H. destructor. Resistant populations were segregating for G119S, A201S, F331Y at the canonical ace. A subset of populations also had copy numbers of canonical ace > 2, which potentially helps overexpress proteins carrying these target-site mutations. Haplotypes possessing different copy numbers and target-site mutations of the canonical ace gene may be under selection across H. destructor populations. We also detected some evidence that increases in copy number of radiated ace-like genes are associated with organophosphate insensitivity, which might suggest potential roles in sequestration or breakdown of organophosphates.

Conclusion

Different combinations of target-site mutations and (or) copy number variation in the canonical ace and ace-like genes may provide non-convergent ways for H. destructor to respond to organophosphate selection. However, these changes may only play a partial role in organophosphate insensitivity, which appears to have a polygenic architecture. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

Comparative laboratory evaluation of the acute and chronic toxicology of diflubenzuron,hexaflumuron and teflubenzuron against II instar desert locust (Schistocerca gregaria) (Orthoptera: Acrididae)

The acute and chronic toxicities of three benzoylphenyl ureas, diflubenzuron, hexaflumuron and teflubenzuron were assessed under laboratory conditions against two-day-old second (II) instar Desert Locust, Schistocerca gregaria (ForskÅl) nymphs. Following exposure by ingestion of a single precise dose applied to short pieces of spring barley, nymphs were monitored for two moults until the fourth (IV) instar. Analysis of acute response data gave three significantly different LD₅₀ statistics (P < 0·05), 68·0, 26·6 and 0·71 μg per nymph respectively for diflubenzuron, hexaflumuron and teflubenzuron. The probit regression slopes also differed significantly, indicating distinct tolerance distributions for the three compounds, the narrowest response being to diflubenzuron and the widest range of response being to teflubenzuron. The timing of death was found to vary between the compounds; most nymphs died during the first moult following treatment with either hexaflumuron or teflubenzuron. However, the majority of nymphs that died after exposure to diflubenzuron did so after completing the first moult after treatment, but before the second. The mean development times of nymphs during the II and especially third (III) instars were significantly longer (P < 0·05) than those of the controls following exposure to diflubenzuron and hexaflumuron. Teflubenzuron had no significant effect (P < 0·05) on the duration of the II instar. The potential of the three compounds to control S. gregaria populations in the field is discussed with particular reference to the timing and nature of acute and chronic responses.     

Foliar lifespan,phenology and seasonal dynamics of the invasive shrub Schinus terebinthifolia

Schinus terebinthifolia is a dioecious tree native to South America that has become an invasive weed in Florida, southern California, southern Arizona, Texas and Hawaii and has been naturalised in over 20 countries. Biological control is considered a viable long-term control option for S. terebinthifolia because release from natural enemies appears to be at least partly responsible for its success in Florida. We examined leaf phenology of S. terebinthifolia over a period of 15 months at five sites in central and southern Florida to provide information that may help in predicting the impacts of potential biocontrol agents for this weed. We documented leaf lifespan, the seasonality of leaf development and abscission and the survivorship of leaves that emerged during either spring, summer or autumn. Average leaf lifespan was >4.5 months at all sites, and leaf phenology followed the seasons closely. Although S. terebinthifolia possesses leaves throughout the year, leaf production was greatest from April to September, and most leaves were abscised in February and March. Spring- and summer-emerging leaves were also longer-lived than leaves produced during autumn. These results suggest that leaves of S. terebinthifolia would be most vulnerable to herbivory during the spring and summer months when newly growing leaf tissue is most plentiful. Biocontrol agents capable of damaging these tissues during spring/summer might be an effective means of controlling this invasive weed.     

Improved detection of Mycobacterium tuberculosis and M. bovis in African wildlife samples using cationic peptide decontamination and mycobacterial culture supplementation

In South Africa, mycobacterial culture is regarded as the gold standard for the detection of Mycobacterium tuberculosis complex (MTBC) infection in wildlife even though it is regarded as “imperfect.” We compared a novel decontamination and mycobacterial culture technique (TiKa) to the conventional mycobacterium growth indicator tube (MGIT) system using known amounts of bacilli and clinical samples from MTBC-infected African buffaloes (Syncerus caffer), white rhinoceros (Ceratotherium simum), and African elephants (Loxodonta africana). Use of the TiKa-KiC decontamination agent on samples spiked with 10,000 to 10 colony forming units (cfu) of M. bovis (SB0121) and M. tuberculosis (H37Rv) had no effect on isolate recovery in culture. In contrast, decontamination with MGIT MycoPrep resulted in no growth of M. bovis samples at concentrations < 1,000 cfu and M. tuberculosis samples < 100 cfu. Subsequently, we used the TiKa system with stored clinical samples (various lymphatic tissues) collected from wildlife and paucibacillary bronchoalveolar lavage fluid, trunk washes, and endotracheal tube washes from 3 species with known MTBC infections. Overall, MTBC recovery by culture was improved significantly (p < 0.01) by using TiKa compared to conventional MGIT, with 54 of 57 positive specimens versus 25 of 57 positive specimens, respectively. The TiKa mycobacterial growth system appears to significantly enhance the recovery of MTBC members from tissue and paucibacillary respiratory samples collected from African buffaloes, African elephants, and white rhinoceros. Moreover, the TiKa system may improve success of MTBC culture from various sample types previously deemed unculturable from other species.