Tuberculosis in wild seals and characterisation of the seal bacillus

SUMMARY Tuberculosis was diagnosed in 3 otariid seals found dead on beaches at 3 locations on the south coast of Western Australian between May 1990 and March 1991. This confirms that tuberculosis is present in the 2 native seals (Neophoca cinerea and Arctocephalus forsteri) in Western Australian waters. Mycobacterium sp isolated from the lungs of 2 of the seals were studied to determine the similarity of the strains to each other, to the strains isolated during 1986 from Australian sea lions and New Zealand fur seals kept in captivity at a marine park near Perth, Western Australia, and to a strain isolated in 1988 from a seal trainer who worked with the infected captive seals for 3 years. After restriction endonuclease analysis (REA) with the endonucleases Bst Ell, Bcl I and Pvu II, one of the wild seal strains appeared to have identical DNA fragment patterns to the strains from the captive seals and the seal trainer. The other wild seal isolate had identical REA profiles using Bst EII and Bcl I, but a minor difference was detected using Pvu II. Differences in these isolates were more clearly seen in restriction fragment length polymorphisms after hybridisation with two DNA probes. The secretory protein MPB70, present in M bovis, was not detected in wild seal isolates using sodium dodecyl sulphate polyacrylamide gel electrophoresis and Western blotting techniques. Analysis of protein and DNA fragment profiles indicated that seal tuberculosis isolates form a unique cluster within the M tuberculosis complex.     

Investigation of Cervical Patency and Uterine Appearance in Domestic Cats by Fluoroscopy and Scintigraphy

The cervical patency of six domestic female cats was monitored under sedation by infusion of contrast medium (Omnipaque) into the cranial vagina during early oestrus, mid‐oestrus, late oestrus and interoestrus or a radiopharmaceutical (^99mTc‐HSA) during mid‐ and interoestrus in a non‐ovulatory oestrous cycle. The transport of the contrast medium or the radiopharmaceutical through the cervix and within the uterine horns was observed under fluoroscopy and with the aid of scintigraphy. In three of the queens, transcervical transport of contrast medium was demonstrated in all stages of oestrus, in one queen during mid‐oestrus, late oestrus and 1 day after oestrus, and in two queens only during late oestrus. The relations between the cervical patency to the contrast medium and the oestrous behaviour, cornification of the vaginal cells and the serum oestradiol‐17β concentration were evaluated, and a relationship was found between the cervical patency and the degree of vaginal cornification. Transcervical transport of the radiopharmaceutical was observed in three queens during mid‐oestrus. When the cervix was open, hysterography under a fluoroscope and hysteroscintigraphy were performed. The fluoroscopic and scintigraphic recordings revealed the patterns of the uterine contractions during oestrus in both ascending and descending directions, and the movement of the uterine contents back and forth between the uterine horns. The hysterograms were classified according to the shape of the uterine horns and the appearance of the endometrial lining. Spiral‐shaped uterine horns with a smooth inner contour were observed in two queens, and a corkscrew appearance with irregular filling defects in the uterine lumen was shown in two queens that had developed subclinical cystic endometrial hyperplasia. These findings demonstrated that fluids or particles deposited in the cranial vagina of the cat can be transported into the uterus during some stages of the oestrous cycle. The fluoroscopic and scintigraphic techniques developed in this study may be further modified to permit more detailed studies of uterine contractile patterns and sperm transport in the feline female reproductive tract. Hysterography proved useful to diagnose uterine disease. The information on cervical patency is of value also for the development of techniques for artificial insemination in this species, and should be studied also in the ovulatory cycle.     

Pathogenesis of placentitis in the goat inoculated with Brucella abortus. II. Ultrastructural studies

Pregnant goats were inoculated intravenously or in uterine arteries with Brucella abortus, and tissues from the uterus and placenta were examined by electron microscopy. Identification of B. abortus in placentae was with antibody-coated colloidal gold. B. abortus was first seen in phagosomes of erythrophagocytic trophoblasts and in the rough endoplasmic reticulum of chorioallantoic trophoblasts. Subsequently, trophoblast necrosis and ulceration of chorioallantoic membranes were present. Coincidently, B. abortus was present in the lumen of placental capillaries. In late stages of infection, placental vasculitis was present, and placentomal trophoblasts were separated from maternal syncytial epithelium. In lesions with vasculitis, large numbers of B. abortus were in connective tissue of chorionic villi. Within the placentome, trophoblasts that lined chorionic villi contained no intracellular bacteria and were separated from B. abortus by intact basement membranes. These results suggest that bacteremic B. abortus is endocytosed by erythrophagocytic trophoblasts and that B. abortus replicates in the rough endoplasmic reticulum of chorioallantoic trophoblasts. Replication of brucellae in trophoblastic rough endoplasmic reticulum is unique; we believe that B. abortus may utilize endoplasmic reticulum for synthesis and glycosylation of bacterial membrane proteins or that B. abortus catabolizes trophoblast secretory proteins.     

Long-term application of animal slurries to grassland alters soil cation balance

Abstract. Soil samples from a 32-year grassland field experiment were taken from 0–5, 5–10, and 10–15 cm soil depths in February 2002. Plots received annual treatments of unamended control, mineral fertilizer, three rates of pig slurry and three rates of cow slurry, each with six replicates. Samples were analysed for cation exchange capacity (CEC), exchangeable cations (Na⁺, K⁺, Ca²⁺, Mg²⁺), pH and Olsen P. Exchangeable sodium percentage (ESP) was calculated as a sodicity indicator. Mean ESP was generally greater for slurry treatments than the control, with a trend of increasing ESP with application rate. This was particularly marked for cow slurry. At 0–5 cm depth ESP increased from 1.18 in the control to 1.75 at the highest rate of pig slurry and 5.60 at the highest rate of cow slurry. Similar trends were shown for CEC, exchangeable Na⁺, K⁺ and Mg²⁺, Ca²⁺ and Olsen P. The build-up of soil P due to slurry applications, together with this combination of physical and chemical factors, may increase the risk of P loss to surface waters, particularly from soils receiving high rates of cow slurry.     

Foxtail-induced ulcerative stomatitis outbreak in a Missouri stable.

Twenty of 25 horses in a well-managed Missouri boarding stable were diagnosed with gingivitis/stomatitis. Gross examination of the affected horses revealed varying degrees of gingivitis ranging from mild periodontal swelling to marked swelling and erythema with ulceration and hemorrhage. Fine hair-like material was embedded within the intensely affected areas. Gingival biopsies from 4 affected horses contained pyogranulomatous inflammation with, in some cases, numerous eosinophils and several grass awns in cross and longitudinal section. Numerous foxtail seed heads were identified in hay samples. Examination of the records revealed that all of the affected horses had been fed the suspect hay, with the exception of 1 horse. Although not deliberately fed the suspect hay, this horse did have access to the hay when turned out into the exercise paddock. The lesions resolved following a change in hay source.     

Physiological Basis of Irrigation Scheduling for Seed Production in Egyptian Clover Syn. Berseem (Trifolium alexandrinum L.) Crop

A field experiment was conducted to find out the critical physiological stages of irrigation schedules inducing better growth, physiological efficiency and seed yield potential of berseem ( Trifolium alexandrinum L., Var. S-99-1). For this purpose eight irrigation treatments were made comprised of four treatments of three irrigation (W₁, W₂, W₃ and W₄), three treatments of four irrigation (W₅, W₇ and W₈) and one treatment of five irrigation (W₆) at various physiological stages i. e. regeneration, flower initiation, full bloom, seed initiation and advance seed development stage.
Thus based on the experimental results the physiological role of watering in berseem seed production could be discussed as:
With-holding of irrigation either at regeneration or at full bloom stage developed potential water stress in plants as indicated by high proline content of irrigation treatments — W₄, W₁ and W₇; and further brought out disturbance on the formation of carotene, synthesis of water soluble sugar and translocation of sugar towards reproductive organs during grain development stage. These stresses adversely affected the plant growth and flowering behaviour. The irrigation at seed initiation stage increased the seed yield. Continuous irrigation did not appear to be useful. Thus it can be concluded that irrigation at three critical physiological stages i. e. regeneration, full bloom and seed initiation was found to be essential for obtaining potential seed yield of berseem.     

Localization of extracellular matrix receptors in ICGN mice, a strain of mice with hereditary nephrotic syndrome.

Fibrotic degeneration was examined in the kidneys of ICR-derived glomerulonephritis (ICGN) mice, a novel inbred mouse line with a hereditary nephrotic syndrome of unknown etiology considered to be a good model of human idiopathic nephrotic syndrome. In the present study, we histochemically revealed changes in accumulation of extracellular matrix (ECM) components and in localization of integrins, cellular receptors for ECM, in the kidneys of ICGN mice with the progression of renal failure. Excessive accumulation of basement membrane (laminin and collagen IV) and interstitial (type III collagen) ECM components were demonstrated in the glomeruli and tubulointerstitum of ICGN mice. Marked deposition of type I collagen and tenascin was seen only in the glomeruli of ICGN mice but not in those of ICR mice as normal controls. Increased expression of integrin alpha1-, alpha2-, alpha5- and beta1-subunits in glomeruli with fibrotic degeneration and abnormal distribution of alpha6-subunit were noted in the kidneys of ICGN mice. Excessive laminin, a ligand of alpha6beta1-integrin, was demonstrated on the tubular basement membrane, but alpha6-subunit diffusely disappeared on the basal side of the tubular epithelial cells. We presumed that abnormal integrin expression in renal tubules causes epithelial cell detachment, and consequently tubular nephropathy, and results in disorder of ECM metabolism causing excessive accumulation of ECM components in the kidneys of ICGN mice.     

A novel approach to studying the effects of temperature on soil biogeochemistry using a thermal gradient bar

The temperature dependence of chemical reaction rates and microbial metabolism mean that temperature is a key factor regulating soil trace gas emissions and hydrochemistry. Here we evaluated a novel approach for studying the thermal response of soils, by examining the effects of temperature on gas emissions and hydrochemistry in (a) peat and (b) soil from a Sitka spruce plantation. A thermal gradient was applied along an aluminium bar, allowing soil to be incubated contemporaneously from 2 to 18 °C. The approach demonstrated clear differences in the biogeochemical responses of the two soil types to warming. The peat showed no significant emission of CH₄ at temperatures below 6 °C, while above 6 °C, a marked increase in the rate of release was apparent up to 15 °C (Q₁₀ = 2.5) with emissions being similar between 15 and 18 °C. Conversely, CH₄ emissions from the forest soil did not respond to warming. Nitrate availability in the peat decreased by 90% between 2 and 18 °C (P < 0.01), whereas concentrations in the forest soil did not respond. Sulphate availability in the peat decreased significantly with warming (60%, P < 0.01), while the forest soil showed the opposite response (a 30% increase, P < 0.01). Conventionally, thermal responses are studied by incubating individual soil samples at different temperatures, involving lengthy preparation and facilities to incubate samples at different temperatures simultaneously. Data collected on a given thermal response is usually limited and thus interpolated or extrapolated. The thermal gradient method overcomes these problems, is simple and flexible, and can be adapted for a wide range of sample types (not confined to soil). Such apparatus may prove useful in the optimization of management practices to mitigate the effects of climate change, as thermal responses will differ depending on land use and soil type.     

Characteristics of cytotoxic cells induced by Toxoplasma lysate antigen in mouse spleen.

Spleen cells from Toxoplasma lysate antigen (TLA)-sensitized BALB/c mice showed the strong cytotoxic activity against both natural killer (NK)-sensitive cells (YAC-1 and RL male-1) and NK-insensitive cells (P-815), when incubated with TLA or recombinant human IL-2 (rhIL-2). The increment of TLA concentration in culture medium increased the cytotoxic activity. Treatment of effector cells; spleen cells from TLA-sensitized mice incubated with TLA, with anti-asialo GM1 or anti-Thy-1 plus complement inhibited the cytotoxic activity of effector cells, whereas treatment with anti-mouse Lyt-2.2 serum plus complement had no effect on the cytotoxic activity. Treatment of spleen cells from TLA-sensitized mice with anti-asialo GM1 and/or anti-Thy-1 plus complement inhibited cytotoxic activities of effector cells. These results suggested that spleen cells sensitized with TLA both in vivo and in vitro were asialo GM1 positive and Thy-1 positive, and the majority of cytotoxic cells induced by TLA were similar to lymphokine-activated killer (LAK) cells induced by IL-2.