Identification of Stably Expressed Reference Genes for RT‐qPCR Data Normalization in Defined Localizations of Cyclic Bovine Ovaries

Ovaries are highly complex organs displaying morphological, molecular and functional differences between their cortical zona parenchymatosa and medullary zona vasculosa, and also between the different cyclic luteal stages. Objective of the present study was to validate expression stability of twelve putative reference genes (RGs) in bovine ovaries, considering the intrinsic heterogeneity of bovine ovarian tissue with regard to different luteal stages and intra‐ovarian localizations. The focus was on identifying RGs, which are suitable to normalize RT‐qPCR results of ovaries collected from clinical healthy cattle, irrespective of localization and the hormonal stage. Expression profiles of twelve potential reference genes (GAPDH, ACTB, YWHAZ, HPRT1, SDHA, UBA52, POLR2C, RPS9, ACTG2, H3F3B, RPS18 and RPL19) were analysed. Evaluation of gene expression differences was performed using genorm , normfinder , and bestkeeper software. The most stably expressed genes according to genorm , normfinder and bestkeeper approaches contained the candidates H3F3B, RPS9, YWHAZ, RPS18, POLR2C and UBA52. Of this group, the genes YWHAZ, H3F3B and RPS9 could be recommended as best‐suited RGs for normalization purposes on healthy bovine ovaries irrespective of the luteal stage or intra‐ovarian localization.     

Larval development of Sesarma catenate Ortmann (Brachyura,Grapsidae, Sesarminae) reared in the laboratory

Sesarma catenata larvae were raised in the laboratory and the development stages described in detail. Larval development comprises four zoea and one megalopa stage. Morphological features of the first zoea and megalopa stages of Sesarma catenata are compared with those of other Sesarminae species described in the literature. The genera Sesarma, Metasesarma, and Bresedium appear to be closely related taxa within the subfamily. The diagnostic value of some features of larval morphology is briefly discussed.     

A comparative analysis of the morphology and nutritive value of five South African native grass species grown under controlled conditions

The comparative growth habits and nutritive value of native grass species of South Africa are largely unknown despite the utility of this information in rangeland restoration efforts. This article presents a comparative characterisation of the morphology, chemical composition and in vitro ruminal fermentation of Urochloa mosambicensis, Cymbopogon pospischilii, Eragrostis superba, Fingerhuthia africana and Eragrostis bicolor when grown under controlled conditions. Species were analysed for height, number of leaves, number of tillers, stem diameter and leaf width at different growth stages, whereas chemical composition and in vitro ruminal dry matter degradability (DMD) were assessed at maturity stage. Grass species and growth stages significantly influenced morphological characteristics. Eragrostis superba and U. mosambicensis had the highest number of leaves at the reproductive stage. Urochloa mosambicensis ranked highest in terms of rangeland restoration potential when all morphological parameters were considered, followed by E. superba and C. pospischilii. With the highest crude protein, low acid-detergent lignin and higher DMD at 48 h, F. africana and C. pospischilii have the highest potential feed value. Across the three most suitable native grass species, U. mosambicensis, E. superba and C. pospischilii, there is sufficient genetic diversity that suggests that these plants may play different and complementary ecological roles in the communal rangeland ecosystem.     

Role of Low-Molecular-Weight Ciliostatic Toxins in Vibriosis of Bivalve Mollusks

Abstract

Toxins play a major role in bacillary necrosis of hatchery-reared bivalve larvae. Two principal types of toxin of Vibrio spp. have been identified: A proteinase (molecular weight = 40,000) that degrades connective tissue and at least one ciliostatic toxin (molecular weight = 500–1,000). The ciliostatic toxin is stable at 100°C for 10 min and probably accounts for the cessation of feeding and swimming in the early stages of bacillary necrosis. Ciliostatic toxins were produced by 17 of 20 Vibrio spp. that are pathogenic to fish or that were isolated from diseased mollusks and included Vibrio tubiashii, V. anguillarum, and V. alginolyticus. Of 53 Vibrio and Aeromonas isolates not associated with bivalve infections, only 15 produced a ciliostatic toxin.     

Pseudorhabdosynochus kritskyi n. sp. (Monogenea: Diplectanidae) on Gag from the Gulf of Mexico

Abstract

Pseudorhabdosynochus kritskyi n. sp. from gag Mycteroperca microlepis (Perciformes: Serranidae) of the Gulf of Mexico is described and differs from all known species of the genus by the shape of the vagina. It may be further differentiated from P. americanum, P. amplidiscatum, P. caballeroi, P. epinepheli, P. latesi, P. monaensis, and Cycloplectanum magnisquamodiscum in having fewer rows of scales in each squamodisc. This new species is most similar to P. beverleyburtonae, P. bocquetae, P. cupatum, P. lantauensis, P. melanesiensis, P. querni, P. serrani, P. summanae, P. vagampullum, and Cycloplectanum riouxi in having 15 or fewer squamodisc rows, but it may be separated by differences in either squamodisc morphology or differently shaped hamuli or bars.     

Sarcocystid organisms found in bile from a dog with acute hepatitis: a case report and review of intestinal and hepatobiliary Sarcocystidae infections in dogs and cats

Sarcocystidae is a family of coccidian protozoa from the phylum Apicomplexa that includes Toxoplasma, Neospora, Sarcocystis, Hammondia, and Besnoitia spp. All species undergo a 2‐host sexual and asexual cycle. In the definitive host, replication is enteroepithelial, and infection is typically asymptomatic or less commonly causes mild diarrhea. Clinical disease is most frequently observed in the intermediate host, often as an aberrant infection, and is mostly associated with neurologic, muscular, or hepatic inflammation. Here, we review the literature regarding intestinal Sarcocystidae infections in dogs and cats, with emphasis on the life cycle stages and the available diagnostic assays and their limitations. We also report the diagnostic findings for an 11‐year‐old dog with acute neutrophilic hepatitis, biliary protozoa, and negative biliary culture. Although Toxoplasma and Neospora IgG titers were both high, PCR for these 2 organisms was negative for bile. The organisms were identified by 18S rDNA PCR as most consistent with Hammondia, either H heydorni or H triffittae. This is the first report of presumed Hammondia organisms being found in canine bile.     

Observations on the Plerocercoid Stage of the Tapeworm Ligula in Three Species of Fish from the Lower Crooked River of Central Oregon

Abstract

Tapeworms in the genus Ligula are cosmopolitan parasites whose plerocercoid stages are found worldwide in fishes of the families Cyprinidae and Catostomidae. Examination of chiselmouth Acrocheilus alutaceus, northern pikeminnow Ptychocheilus oregonensis, and bridgelip sucker Catostomus columbianus revealed the presence of Ligula in the lower Crooked River system of central Oregon. This represents the first report of Ligula intestinalis in two of these host species: chiselmouth and bridgelip sucker. Although most hosts appeared healthy, large external lesions were noted on some bridgelip suckers in close association with a Ligula plerocercoid. The largest plerocercoid recovered was 101.5 cm relaxed length and may represent the longest plerocercoid of Ligula reported from North America.     

Co-culture model reveals the characteristics of theca cells and the effect of granulosa cells on theca cells at different stages of follicular development

Theca cells (TCs) play an important role in follicular development, which cannot be separated from granulosa cells (GCs). However, compared with mammals, the TCs and the effects of GCs on TCs at different follicular development stages (FDSs) have specific characteristics in avian species, but none of them have been clearly defined. In this study, we established an in vitro co-culture (with GC at the corresponding stage) model of goose TCs at different FDSs (pre-hierarchical, hierarchical and F1) by using a transwell system. The properties of TCs in co-culture at the three FDSs, including cell morphology, activity and intracellular lipid content, as well as the expression of key genes involved in de novo lipogenesis, steroidogenesis, proliferation and apoptosis, were examined and defined. We further compared the mono-culture and co-culture groups. After co-culture, the activity of TCs showed significant (p < .01) increases in all stages; moreover, in pre-hierarchical TCs, the expression levels of FAS, SREBP, 3β-HSD and CCND1 were promoted, and PPARγ, CYP19, BCL2 and CAS3 were inhibited (p < .05); in the hierarchical TCs, the expression levels of PPARγ, FAS, CYP19, CCND1 and BCL2 were promoted, and SREBP, STAR, 3β-HSD and CAS3 were inhibited (p < .05), whereas in the F1 TCs, the expression levels of PPARγ, FAS, 3β-HSD, CYP19 and CCND1 were promoted, and STAR and CAS3 were inhibited (p < .05). These results suggested that GCs at the three FDSs have dynamic and complex influences on the physiological characteristics of TCs, and the influences on TCs at the three FDSs were varied.     

Bacterial, fungal and parasitic infections in the ostrich (Struthio camelus var. domesticus)

The ostrich is susceptible to microorganisms of bacterial, fungal and parasitic origin. Anthrax, caused by Bacillus anthracis, is dangerous to other livestock and humans. Salmonella is transmitted from rodents or wild bird reservoirs. Pausterellosis caused by Pasteurella multocida results in air sac infections in ostriches. Colibacillosis is caused by Escherichia coli. Tuberculosis caused by Mycobacterium avium, is very rare in ostriches. Aspergillosis principally afflicts chicks. Zygomycosis, a secondary fungal infection of the upper gastrointestinal tract, is caused by Basidia, Mucor and Rhizopus. Leucocytozoon struthionis and Plasmodium spp. are harmless protozoa transmitted from flying arthropods. The tapeworm, Houttuynia struthionis, is dangerous in young ostriches. The adult ratite fluke (Philophthalmus gralli) is transmitted to ostriches following ingestion of infected freshwater crustaceans. Tick infestations of ostrich skin in Africa include Amblyomma spp., Haemaphysalis punctata, Hyalomma spp., Rhipicephalus turanicus and Argus spp. The ostrich quillmite (Pterolichus bicaudatus) and louse (Struthioliperus struthionus) may lower skin and leather quality via pruritis and/or excessive preening and feather loss. Nemododa infections are rare.     

Ultra‐frequent HRAS p.Q61R somatic mutation in canine acanthomatous ameloblastoma reveals pathogenic similarities with human ameloblastoma

Ameloblastoma is a locally aggressive odontogenic tumour that occurs in humans and dogs. Most ameloblastomas (AM) in humans harbour mutually‐exclusive driving mutations in BRAF, HRAS, KRAS, NRAS or FGFR2 that activate MAPK signalling, and in SMO that activates Hedgehog signalling. The remarkable clinical and histological similarities between canine acanthomatous ameloblastoma (CAA) and AM suggest they may harbour similar driving mutations. In this study, aimed at characterizing the mutational status of SMO, BRAF, HRAS, KRAS, NRAS and FGFR2 in CAA, we used RNA sequencing, Sanger sequencing and restriction fragment length polymorphism assays to demonstrate that 94% of CAA (n = 16) harbour a somatic HRAS p.Q61R mutation. The similarities in MAPK‐activating mutational profiles between CAA and AM implicate conserved molecular mechanisms of tumorigenesis, thus, qualifying the dog as a potentially useful model of disease. Given the relevance of RAS mutations in the pathogenesis of odontogenic tumours and other types of cancer, the results of this study are of comparative, translational, and veterinary value.     

Early development and larval behaviour of a minnow,Barbus anoplus (Pisces: Cyprinidae)

The chubbyhead barb, Barbus anoplus, underwent a population explosion in the early phases of filling of Lake be Roux on the Orange River. This successful colonization was possibly related to the survival strategy of the young stages of this minnow. It is suggested that some of the development traits of B. anoplus enabled it to become the most widespread freshwater fish species south of the Limpopo River. The development and behaviour of embryos and larvae of B. anoplus are described and discussed with reference to their survival strategy and potential colonizing ability. Some of the protolarvae were pelagic and the relevance of this behaviour is noted.     

Dose confirmation studies for monepantel, an amino-acetonitrile derivative, against fourth stage gastro-intestinal nematode larvae infecting sheep

Monepantel is the first compound from the recently discovered amino-acetonitrile derivative (AAD) class of anthelmintics to be developed for use in sheep. Nine dose confirmation studies were conducted in Australia, New Zealand and Switzerland to confirm the minimum therapeutic oral dose of monepantel to control fourth stage (L4) gastro-intestinal nematode larvae in sheep (target species were Haemonchus contortus, Teladorsagia (Ostertagia) circumcincta, Teladorsagia trifurcata, Trichostrongylus axei, Trichostrongylus colubriformis, Trichostrongylus vitrinus, Cooperia curticei, Cooperia oncophora, Nematodirus battus, Nematodirus filicollis, Nematodirus spathiger, Chabertia ovina and Oesophagostomum venulosum). In each study, sheep infected with a defined selection of the target nematodes were treated with 2.5 mg monepantel/kg liveweight. Following euthanasia and worm counting, efficacy was calculated against worm counts from untreated control groups. The results demonstrate high (95 < 100%) efficacy of monepantel when administered orally to sheep at 2.5 mg/kg for most species tested. Efficacy levels against N. spathiger and O. venulosum were variable and failed to meet the required regulatory standard (≥90%) in some studies. Efficacy was demonstrated against L4 stages of nematodes known to be resistant to either benzimidazole and/or levamisole anthelmintics (macrocyclic lactone resistant isolates were not available for testing). The broad-spectrum activity of monepantel against L4 larvae of common gastro-intestinal nematodes in sheep and its favorable safety profile represents a significant advance in the treatment of parasitic gastro-enteritis in this animal species. No adverse effects related to treatment with monepantel were observed.     

The efficacy of oxfendazole against natural infections of nematodes & cestodes in sheep

Oxfendazole (5 mg/kg) was tested against natural infections of nematodes and cestodes in sheep in three experiments.

It was 100% effective against all stages of Ostertagia sp., Nematodirus spathiger, Oesophagostomum venulosum, Chabertia ovina, Moniezia expansa, immature 5th stage and adult Cooperia curticei and 4th stage Trichostrongylus sp. Efficacy was 99%–100% against immature 5th stage and adult Trichostrongylus sp., 67.8%and 91.6% against adult Strongyloides sp., 80.7%and 89.5% against immature 5th stage Trichuris ovis and 74% and 79% against Trichuris ovis.     

Hatchery Efficacy Trials with Chloramine-T for Control of Bacterial Gill Disease

Abstract

Three hatchery trials were carried out to determine the efficacy of chloramine-T for the control of bacterial gill disease in rainbow trout Oncorhynchus mykiss, ranging in weight from 2.5 to 54 g. In each trial, we used a single flow-through treatment of 8.5 mg chloramine-T/L. The single treatment provided effective control in all three trials; however, results were best when treatment was begun in the early stages of an outbreak. A second or third treatment may be required if an outbreak is in an advanced stage or if fish are under stress.