Isolation of a selected microbial consortium from a pesticide-contaminated mix-load site soil capable of degrading the herbicides atrazine and alachlor

Contaminated soil from a 100-year-old mix-load site located in Reading, PA was evaluated for its potential to provide indigenous microorganisms capable of degrading two widely utilized herbicides, atrazine (2-chloro-4-ethylamino-6-isopropylamino-S-triazine; AT) and alachlor (2-chloro-2′,6′-diethyl-N-[methoxymethyl]-acetanilide; AL). Three different locations from the site were chosen for experimentation based on herbicide handling activities. Standard enrichment techniques were used to isolate a selective microbial consortium (SCM) with the desirable degrading capabilities. Three enrichment treatment schemes were evaluated; AT and AL, AL alone, and only AT. Degradative organisms were isolated from only one of the sample locations. Considerable differences in the soil parameters of the three sample locations were found that might have had an effect on the ability of the indigenous microbial populations within the soil to degrade AT and AL. In the initial cultures from this location, degradation occurred in the AT and AL treatment only. Because the AT and AL were the only sources of carbon and nitrogen (N) for the microbes, these results suggest that AL alone was not a sufficient N source. In general, the ability to degrade AL by the SMC was dependent on AT degradation. Alachlor degradation did not begin until approximately 15% of the AT was transformed. Once all of the AT was removed very little further AL degradation occurred. The average half-life (t_1/2) of AT was 7.5 d, while average t_1/2 for AL degradation was 11 d. Individual colonies from the SMC were identified by fatty acids methyl ester (FAME) analysis. Five strains were identified with similarity indexes above 70%. These isolates included the following: Alcaligenes xylosoxydans subsp. denitrificans, Alcaligenes xylosoxydans subsp. xylosoxydans, Pseudomonas putida, Pseudomonas marginalis, and Providencia rustigianii.     

Improved bioavailability of dietary phenolic acids in whole grain barley and oat groat following fermentation with probiotic Lactobacillus acidophilus , Lactobacillus johnsonii , and Lactobacillus reuteri

The aim of this study was to improve the bioavailability of the dietary phenolic acids in flours from whole grain barley and oat groat following fermentation with lactic acid bacteria (LAB) exhibiting high feruloyl esterase activity (FAE). The highest increase of free phenolic acids was observed after fermentation with three probiotic strains, Lactobacillus johnsonii LA1, Lactobacillus reuteri SD2112, and Lactobacillus acidophilus LA-5, with maximum increases from 2.55 to 69.91 μg g(-1) DM and from 4.13 to 109.42 μg g(-1) DM in whole grain barley and oat groat, respectively. Interestingly, higher amounts of bound phenolic acids were detected after both water treatment and LAB fermentation in whole grain barley, indicating higher bioaccessibility, whereas some decrease was detected in oat groat. To conclude, cereal fermentation with specific probiotic strains can lead to significant increase of free phenolic acids, thereby improving their bioavailability.     

Identification of genomic alterations with clinical impact in canine splenic hemangiosarcoma

Canine hemangiosarcoma (HSA) is an aggressive cancer of endothelial cells with short survival times. Understanding the genomic landscape of HSA may aid in developing therapeutic strategies for dogs and may also inform therapies for the rare and aggressive human cancer angiosarcoma. The objectives of this study were to build a framework for leveraging real-world genomic and clinical data that could provide the foundation for precision medicine in veterinary oncology, and to determine the relationships between genomic and clinical features in canine splenic HSA. One hundred and nine dogs with primary splenic HSA treated by splenectomy that had tumour sequencing via the FidoCure® Precision Medicine Platform targeted sequencing panel were enrolled. Patient signalment, weight, metastasis at diagnosis and overall survival time were retrospectively evaluated. The incidence of genomic alterations in individual genes and their relationship to patient variables including outcome were assessed. Somatic mutations in TP53 (n = 44), NRAS (n = 20) and PIK3CA (n = 19) were most common. Survival was associated with presence of metastases at diagnosis and germline variants in SETD2 and NOTCH1. Age at diagnosis was associated with somatic NRAS mutations and breed. TP53 and PIK3CA somatic mutations were found in larger dogs, while germline SETD2 variants were found in smaller dogs. We identified both somatic mutations and germline variants associated with clinical variables including age, breed and overall survival. These genetic changes may be useful prognostic factors and provide insight into the genomic landscape of hemangiosarcoma.     

A Collagen Basketweave from the Giant Squid Mantle as a Robust Scaffold for Tissue Engineering

The growing applications of tissue engineering technologies warrant the search and development of biocompatible materials with an appropriate strength and elastic moduli. Here, we have extensively studied a collagenous membrane (GSCM) separated from the mantle of the Giant squid Dosidicus Gigas in order to test its potential applicability in regenerative medicine. To establish the composition and structure of the studied material, we analyzed the GSCM by a variety of techniques, including amino acid analysis, SDS-PAGE, and FTIR. It has been shown that collagen is a main component of the GSCM. The morphology study by different microscopic techniques from nano- to microscale revealed a peculiar packing of collagen fibers forming laminae oriented at 60–90 degrees in respect to each other, which, in turn, formed layers with the thickness of several microns (a basketweave motif). The macro- and micromechanical studies showed high values of the Young’s modulus and tensile strength. No significant cytotoxicity of the studied material was found by the cytotoxicity assay. Thus, the GSCM consists of a reinforced collagen network, has high mechanical characteristics, and is non-toxic, which makes it a good candidate for the creation of a scaffold material for tissue engineering.     

Endocrine and ovarian responses in water buffalo cows immunized against inhibin and subjected to the Ovsynch protocol

The aim of this study was to investigate the feasibility of stimulating ovarian follicle development in order to improve fertility in water buffalo cows by immunization against inhibin. The experiment was carried out in early summer(May) and included 24 multi-parity crossbred Murrah-Swamp buffaloes that were divided into immunized(n=11) and control(n=13) groups. Each immunized cow was administered with a 2-m L immunogen of mineral oil adjuvant containing 2 mg of recombinant inhibin α-subunit fusion protein. The controls were treated with the adjuvant only. All animals received Ovsynch protocol treatment, starting on the day of the antigen administration, and they were artificially inseminated upon behavioral estrus. As a result, all of the immunized buffaloes generated antibodies against inhibin during the experimental period and had higher plasma concentrations of follicle-stimulating hormone(FSH), activin, and estradiol(E2) related to estrous expression. A higher proportion of immunized animals expressed estrus behavior than did the controls(72% vs. 30%, P0.05). On average, inhibin-immunized buffaloes had significantly more large follicles(≥9 mm in diameter) than the controls(mean±SEM; 1.2±0.1 vs. 0.84±0.1, respectively; P0.05) and a slightly higher mean total number of follicles(≥2 mm; 11.4±0.7 vs. 9.0±1.1, respectively; P=0.09) and small(2–4 mm) follicles(8.81±0.6 vs. 6.84±1.0, respectively; P=0.12). A higher percentage of cows ovulated in the immunized group than in the control group(91%(10/11) vs. 54%(7/13), respectively; P0.05). Moreover, inhibin-immunized cows had slightly larger corpus luteum(CL) than the controls 9 days after ovulation and significantly higher(P0.01) post-ovulation peak plasma progesterone(P4) concentrations. Immunization against inhibin also marginally increased the conception rate 42 days after insemination(45.8% vs. 15.4%; P0.05). These results demonstrate that immunization against inhibin, coupled with the treatment with the Ovsynch protocol, can constitute a new technique to increase fertility in water buffalo cows.     

The impacts of forest clearance on lizard, small mammal and bird communities in the arid spiny forest, southern Madagascar

Madagascar is a global biodiversity hotspot threatened by forest loss, degradation and fragmentation, all of which are detrimental to the future survival of forest-dwelling organisms. For conservation purposes it is essential to determine how species respond to habitat disturbance, specifically deforestation. In this study we investigated the impacts of deforestation on three vertebrate communities, lizards, small mammals and birds, in an area of spiny forest subjected to anthropogenic forest clearance. Spiny forest has high levels of endemism, but conservation in this unique ecosystem is hindered by the lack of research. We undertook standardised trapping, time-constrained and timed species searches to assess species richness, species abundance and community composition of lizards, small mammals and birds in six areas of ‘forest’ and six ‘cleared’ areas. From surveys and opportunistic sightings we recorded a total of 70 species of birds, 14 species of mammals and 38 species of reptiles and amphibians. We found forest clearing to have a negative effect on species richness and community structure of all groups and identified loss of canopy cover as a driving factor behind this. However, the response and sensitivity to clearing varied between groups and species. Lizards (50%) and small mammals (40%) had the greatest decline in species richness in response to clearing as compared to birds (26%), although birds showed the greatest shift in community structure. The community in cleared areas contained more generalist and introduced species that have wider geographic ranges and habitat preferences, than those unique to the spiny forest. We found the first suite of species to suffer from forest clearance were those of high conservation priority due to their restricted geographic range. Our findings are discussed in relation to future spiny forest conservation and management.     

Hepatotoxic Seafood Poisoning (HSP) Due to Microcystins: A Threat from the Ocean?

Cyanobacterial blooms are a major and growing problem for freshwater ecosystems worldwide that increasingly concerns public health, with an average of 60% of blooms known to be toxic. The most studied cyanobacterial toxins belong to a family of cyclic heptapeptide hepatotoxins, called microcystins. The microcystins are stable hydrophilic cyclic heptapeptides with a potential to cause cell damage following cellular uptake via organic anion-transporting proteins (OATP). Their intracellular biologic effects presumably involve inhibition of catalytic subunits of protein phosphatases (PP1 and PP2A) and glutathione depletion. The microcystins produced by cyanobacteria pose a serious problem to human health, if they contaminate drinking water or food. These toxins are collectively responsible for human fatalities, as well as continued and widespread poisoning of wild and domestic animals. Although intoxications of aquatic organisms by microcystins have been widely documented for freshwater ecosystems, such poisonings in marine environments have only occasionally been reported. Moreover, these poisonings have been attributed to freshwater cyanobacterial species invading seas of lower salinity (e.g., the Baltic) or to the discharge of freshwater microcystins into the ocean. However, recent data suggest that microcystins are also being produced in the oceans by a number of cosmopolitan marine species, so that Hepatotoxic Seafood Poisoning (HSP) is increasingly recognized as a major health risk that follows consumption of contaminated seafood.     

Lethal Ozolaimus megatyphlon infection in a green iguana (Iguana iguana rhinolopa).

An imported 2.5-yr-old female green iguana (Iguana iguana rhinolopa) kept in Greece was presented with a history of anorexia and allotriophagy of 1 mo duration. Upon clinical examination, it was cachectic and had severe abdominal distension and fibrous osteodystrophy. Despite treatment, it died a month later. On necropsy, massive accumulations of threadlike nematode parasites were observed in the large intestine, identified as Ozolaimus megatyphlon, a member of the Oxyuridae family of Pharyngodonidae, a usually nonpathogenic intestinal parasite of iguanas. To the authors' knowledge, its presence has not been reported previously in Europe, although one pathogenic infection has been reported previously in Japan. The animal was presumably infected before importation. Although death was attributed to the heavy parasitic overload, the poor diet and terrarium hygiene, and absence of an anthelminthic regime further contributed to the deterioration of the animal's condition. Recognition of this condition, which may be recently introduced or underdiagnosed, may help improve medical and trade standards concerning this species in practice.     

Aspergillus fumigatus Bronchopneumonia in a Hellenic Shepherd Dog

A 3 yr old intact female Hellenic shepherd dog was referred due to depression, partial anorexia, fever, and a mild productive cough of 2 mo duration. Thoracic radiographs showed increased opacity of all of the left lung lobes. Upon bronchoscopy, a sanguineous, purulent discharge was detected in the tracheal lumen with hyperplastic tissue narrowing the left main stem bronchus. Cultures were positive for bacteria (Bacillus spp. and Clostridium spp.) but negative for fungi. Due to the severity of the lesions, a complete left lung pneumonectomy was performed. Histopathological examination of the excised lung tissues revealed a severe granulomatous bronchopneumonia with numerous alveolar macrophages laden with structures stained positively by periodic acid-Schiff and Grocott stain that had morphology consistent with fungi. PCR and sequencing of internal transcribed spacer regions 1 and 2 from genetic material extracted from paraffin-embedded pulmonary tissue confirmed the presence of Aspergillus fumigatus. Itraconazole was administrated for 5.5 mo and the dog was clinically normal 26 mo after surgery.     

Coralligenous assemblages along their geographical distribution: Testing of concepts and implications for management

• 1. The coralligenous habitat was studied at the large Mediterranean scale, by applying a standardized, non‐destructive photo‐sampling protocol, developed in the framework of the CIGESMED project.
• 2. The results provided evidence to support the following statements: (a) the assemblage pattern is not homogeneously distributed across the four Mediterranean ecoregions studied (biotic gradients hypothesis); and (b) the assemblage pattern does not change significantly when the information is aggregated to higher taxonomic levels (taxonomic sufficiency hypothesis).
• 3. Surrogate taxonomic categories higher than species, such as genus and family, can be used to reveal the multivariate pattern of the coralligenous assemblages.
• 4. Although preliminary at the pan‐Mediterranean scale, these outcomes set the scene for future comparisons as more data sets become available but also for comparisons between taxonomic and functional patterns.