Ethidium bromide: A general purpose fluorescent stain for nucleic acid in bacteria and eucaryotes and its use in microbial ecology studies     

David J. Roser 《Soil biology & biochemistry》1980,12(4):329-336

The fluorochrome ethidium bromide (EB) combines with double-stranded DNA and fluoresces when excited by u.v. radiation.When applied at aqueous concentrations ranging from 2 mg l^?1 to Ig l^?1 EB strongly stained all eucaryotic and procaryotic cell types tested. It concentrated preferentially in procaryotic cells containing DNA and in the nucleus and other cytoplasmic components of eucaryotic cells. It did not stain cell walls or inert materials, e.g. clay and sand, as intensely as material containing nucleic acid.Potential applications for EB as a stain include the detection of plant root nuclei and the examination of bacteria and fungi in soil and mud preparations.  相似文献   

Pathogenesis and transmissibility of highly (H7N1) and low (H7N9) pathogenic avian influenza virus infection in red-legged partridge (Alectoris rufa)     

Bertran K  Pérez-Ramírez E  Busquets N  Dolz R  Ramis A  Darji A  Abad FX  Valle R  Chaves A  Vergara-Alert J  Barral M  Höfle U  Majó N 《Veterinary research》2011,42(1):24

ABSTRACT: An experimental infection with highly pathogenic avian influenza virus (HPAIV) and low pathogenic avian influenza virus (LPAIV) was carried out in red-legged partridges (Alectoris rufa) in order to study clinical signs, gross and microscopic lesions, and viral distribution in tissues and viral shedding. Birds were infected with a HPAIV subtype H7N1 (A/Chicken/Italy/5093/1999) and a LPAIV subtype H7N9 (A/Anas crecca/Spain/1460/2008). Uninoculated birds were included as contacts in both groups. In HPAIV infected birds, the first clinical signs were observed at 3 dpi, and mortality started at 4 dpi, reaching 100% at 8 dpi. The presence of viral antigen in tissues and viral shedding were confirmed by immunohistochemistry and quantitative real time RT-PCR (qRRT-PCR), respectively, in all birds infected with HPAIV. However, neither clinical signs nor histopathological findings were observed in LPAIV infected partridges. In addition, only short-term viral shedding together with seroconversion was detected in some LPAIV inoculated animals. The present study demonstrates that the red-legged partridge is highly susceptible to the H7N1 HPAIV strain, causing severe disease, mortality and abundant viral shedding and thus contributing to the spread of a potential local outbreak of this virus. In contrast, our results concerning H7N9 LPAIV suggest that the red-legged partridge is not a reservoir species for this virus.  相似文献   

Determination of water-soluble arsenic compounds in commercial edible seaweed by LC-ICPMS     

Llorente-Mirandes T  Ruiz-Chancho MJ  Barbero M  Rubio R  López-Sánchez JF 《Journal of agricultural and food chemistry》2011,59(24):12963-12968

This paper reports arsenic speciation in edible seaweed (from the Galician coast, northwestern Spain) produced for human consumption. Chondrus crispus , Porphyra purpurea , Ulva rigida , Laminaria ochroleuca , Laminaria saccharina , and Undaria pinnatifida were analyzed. The study focused on arsenosugars, the most frequently occurring arsenic species in algae. As(III) and As(V) were also determined in aqueous extracts. Total arsenic in the samples was determined by microwave digestion and inductively coupled plasma mass spectrometry (ICPMS). For arsenic speciation, a water extraction especially suitable for arsenosugars was used, and the arsenic species were analyzed by liquid chromatography with both anionic and cationic exchange and ICPMS detection (LC-ICPMS). The total arsenic content of the alga samples ranged from 5.8 to 56.8 mg As kg(-1). The mass budgets obtained in the extracts (column recovery × extraction efficiency) ranged from 38 to 92% except for U. pinnatifida (4%). The following compounds were detected in the extracts: arsenite (As(III)), arsenate (As(V)), methylarsonate (MA), dimethylarsinate (DMA), sulfonate sugar (SO(3)-sug), phosphate sugar (PO(4)-sug), arsenobetaine (AB), and glycerol sugar (Gly-sug). The highest concentrations corresponded to the arsenosugars.  相似文献   

Testicular function and fertility     

PhDJanet F. Roser 《Journal of Equine Veterinary Science》2000,20(2):90

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Plasma Concentrations of Pregnancy‐Associated Glycoproteins Measured Using Anti‐Bovine PAG‐2 Antibodies on Day 120 of Gestation Predict Abortion in Dairy Cows Naturally Infected with Neospora caninum     

I García‐Ispierto  S Almería  B Serrano  NM de Sousa  JF Beckers  F López‐Gatius 《Reproduction in domestic animals》2013,48(4):613-618

The present study sought to determine: (i) the effects of Neospora caninum infection and twin pregnancy on plasma pregnancy‐associated glycoprotein‐2 (PAG‐2) concentrations throughout pregnancy and (ii) whether plasma PAG‐2 concentrations could predict abortion in N. caninum‐infected cows. The study was performed on a commercial Holstein‐Friesian dairy herd in northeastern Spain and the final data included those recorded in 53 non‐aborting and 19 aborting animals. Blood samples were collected immediately before pregnancy diagnosis (on Days 40, 90, 120, 150, 180 and 210 post‐insemination) in non‐aborting cows or until the time of abortion detection in aborting cows. General lineal models (GLM) repeated measures anova revealed the different behaviour of PAG‐1 and PAG‐2, and significant effects of Neospora seropositivity, cool season and twin pregnancy on plasma PAG‐2 concentrations throughout gestation (between‐subject effects). In addition, based on the odds ratios, the likelihood of abortion increased in Neospora‐seropositive cows (by a factor of 7.0) compared to seronegative animals and decreased in cows with a high plasma PAG‐2 concentration (>4.5 ng/ml) on Day 120 of pregnancy (by a factor of 0.24), compared to the remaining cows. In conclusion, there is a relationship between plasma PAG‐2 concentrations and the risk of abortion in Neospora‐infected dairy cows. Thus, plasma PAG concentrations measured using anti‐boPAG‐2 antiserum on Day 120 of gestation could serve as an indicator of the abortion risk in N. caninum infected animals; values <4.5 ng/ml indicating a high risk of abortion in chronically infected animals.  相似文献   

An International Delphi Study of the Causes of Death and the Criteria Used to Assign Cause of Death in Bovine Perinatal Mortality     

JF Mee  C Sanchez‐Miguel  M Doherty 《Reproduction in domestic animals》2013,48(4):651-659

The objective of the present study was to elicit opinion from two groups of veterinarians [subject matter experts and non‐subject matter experts] about the causes of bovine perinatal mortality and the criteria used to assign such causes. The subject matter experts were selected on the basis of their scientific publications or experience of working in a veterinary diagnostic or research laboratory in the area of bovine perinatal mortality. The non‐subject matter experts were self‐selected as cattle veterinarians without particular expertise in bovine perinatology. A total of 74 veterinarians (46 subject matter experts and 28 non‐subject matter experts) from 23 countries responded. The study was conducted using Delphi methodology over seven rounds. Respondents were asked to agree the causes of bovine perinatal mortality and for each cause to agree the supporting diagnostic criteria. There was a close agreement between groups on 16 causes of death apart from intra‐uterine growth retardation (IUGR) and micronutrient imbalances which were accepted by fewer subject matter experts. There was inter‐group consensus on the criteria to diagnose accidents, congenital defects, dystocia, hyperthermia, infections, premature placental separation, prematurity and prolonged calving. There was inter‐group consensus on the criteria to diagnose anoxia, apart from gingival cyanosis; on haemorrhage, apart from haemorrhagic anaemia; on IUGR, apart from organ weights; and on iodine imbalance, apart from goitre and thyroid iodine content. The results from this study highlighted the current lack of standardization of the criteria used to define the cause of death for bovine perinatal mortality and the need for such standardization.  相似文献   

64, XX,SRY‐negative,Testicular DSD Syndrome in a Lusitano Horse     

A Torres  JF Silva  N Bernardes  J Sales Luís  L Lopes da Costa 《Reproduction in domestic animals》2013,48(3):e33-e37

Here is reported a disorder of sex development found in the Portuguese Lusitano horse breed. The complex genital phenotype included mammary glands, abdominal testes without epididymis, connected through oviducts to pelvic hypoplastic uterine horns and fused vulvar labia majora from which protruded ventrally a penis‐like structure. This structure was presented in a reversed position, the urethral opening placed dorsally in the glans. However, it was functional both for micturition and erection. The horse exhibited female micturition posture and aggressive male‐like behaviour, including flehmen, mounting, thrusting and flagging of the tail. Plasma testosterone concentrations were below detection limits and the genetic evaluation revealed a 64, XX, SRY‐negative karyotype. Surgery consisted in the removal of abdominal gonads followed by amputation of the penis and repositioning of the urethra. This case of reversion between the chromosomal and gonadal sex, associated with mixed anatomical and behavioural phenotype, illustrates that development of the testes may occur in the absence of the SRY gene and that other genetic and cellular pathways leading to gonad differentiation should be investigated.  相似文献   

The occurrence of Cryptosporidium parvum,Campylobacter and Salmonella in newborn dairy calves in the Manawatu region of New Zealand     

A Grinberg  WE Pomroy  JF Weston  A Ayanegui-Alcerreca  D Knight 《New Zealand veterinary journal》2013,61(5):315-320

AIM: To determine the occurrence of Cryptosporidium parvum oocysts, Campylobacter spp and Salmonella spp in faecal samples taken from newborn dairy calves on 24 dairy farms in the Manawatu region of New Zealand.

METHODS: A cross-sectional study was conducted during the 2002 calving season. Faecal samples were collected from 185 newborn calves from a convenience sample of 24 dairy farms. The samples were tested microscopically for the presence of C. parvum oocysts, and bacteriologically for the presence of Campylobacter spp and Salmonella spp.

RESULTS: Infections with C. parvum were identified in 33/156 (21.2%) calves from 10 farms. More than 10⁶ oocysts/g (OPG) faeces were detected in calves from four farms. Campylobacter spp were isolated from 58/161 (36%) calves from 18 farms; in particular, C. jejuni subsp jejuni was isolated from 11/161 (6.8%) calves from seven farms. Salmonellae were not detected.

CONCLUSIONS: Despite the short and concentrated calving pattern and the long interval between calving seasons characterising most dairy farms in New Zealand, C. parvum is widespread among calves. Campylobacter spp, especially C. jejuni, rapidly colonise the intestinal tract of newborn calves.

RELEVANCE: This study provided an estimate of the ecological impact of newborn dairy calves with regard to the potentially zoonotic enteric pathogens most frequently isolated from human gastrointestinal infections in New Zealand.  相似文献   

Pregnancy Rates in Angus Cross Beef Cows Bred at Observed Oestrus With or Without Second GnRH Administration in Fixed‐Time Progesterone‐Supplemented Ovsynch and CO‐Synch Protocols     

R Kasimanickam  JB Hall  JF Currin  B Inman  JS Rudolph  WD Whittier 《Reproduction in domestic animals》2010,45(3):487-492

Crossbred cows (n = 1073) from five locations had oestrous cycles synchronized with 100 μg of GnRH IM and insertion of controlled internal drug release device (CIDR) on Day 0 followed by 25 mg of PGF_2α IM and CIDR removal on Day 7. Kamar^® patches were placed on all cows at CIDR removal. Cows were observed three times daily for oestrus after PGF_2α administration. In the Ovsynch‐CIDR group, cows detected in oestrus (n = 193) within 48 h after PGF_2α were inseminated using the AM–PM rule. Among these cows, 80 received and 113 did not receive a second GnRH at 48 h after PGF_2α. Cows (n = 345) not detected in oestrus received a second GnRH at 48 h after PGF_2α on Day 9, and fixed‐time AI 16 h after the GnRH on Day 10. In the CO‐Synch‐CIDR group, cows detected in oestrus (n = 224) within 48 h after PGF_2α were inseminated using the AM–PM rule. Among these cows, 79 received and 145 did not receive a second GnRH at 64 h after PGF_2α. Cows (n = 311) not detected in oestrus received a second GnRH on Day 10 at the time of AI, 64 h after PGF_2α. The AI pregnancy rates were not different between the Ovsynch‐CIDR and CO‐Synch‐CIDR groups (p = 0.48). There were no differences in the AI pregnancy rates for cows inseminated at a fixed time (p = 0.26) or at detected oestrus (p = 0.79) between the treatment groups. Among cows inseminated in oestrus, there were no differences in the AI pregnancy rates between cows that received or did not receive the second GnRH (p = 0.47). In conclusion, acceptable AI pregnancy rates can be achieved with or without inclusion of oestrus detection in the Ovsynch‐CIDR and CO‐Synch‐CIDR protocols. Among cows detected in oestrus, cows that received a second GnRH yielded similar pregnancy rates when compared with cows that did not receive the second GnRH.  相似文献   

Use of Ox‐Aquaculture© for disinfection of live prey and meagre larvae,Argyrosomus regius (Asso, 1801)          下载免费PDF全文

Roser Vallés  Ana Roque  Ana Caballero  Alicia Estévez 《Aquaculture Research》2015,46(2):413-419

Live prey used for marine larval fish (rotifers and Artemia) as well as intensive larval rearing conditions are susceptible to the proliferation of bacteria that are the cause for reduced growth and larval mortality. Hydrogen peroxide has been recently proved a good disinfectant in aquaculture, either for eggs, larvae or live prey. In this study the effects of a hydrogen peroxide‐based product, Ox‐Aquaculture^©, on live prey (rotifers and Artemia) and meagre larvae bacterial load, composition and final status have been tested. A 34.6% reduction of total heterotrophic bacteria and 59.7% of Vibrionaceae were obtained when rotifers were exposed for 15 min to 40 mg L^?1 of the product. A 34.3% reduction of total heterotrophic bacteria and 37.7% of Vibrionaceae were obtained when Artemia were exposed for 5 min to 8000 mg L^?1 of the product. More than 95% reduction of total heterotrophic bacteria and 75% of Vibrionaceae were obtained when meagre larvae were exposed for 1 h to 20 mg L^?1 of the product. Furthermore, disinfection of enriched live prey with the product did not change the fatty acid composition and survival of the live prey and improved final larval survival.  相似文献