Fungal Planet description sheets: 1112-1181.

Novel species of fungi described in this study include those from various countries as follows: Australia, Austroboletus asper on soil, Cylindromonium alloxyli on leaves of Alloxylon pinnatum, Davidhawksworthia quintiniae on leaves of Quintinia sieberi, Exophiala prostantherae on leaves of Prostanthera sp., Lactifluus lactiglaucus on soil, Linteromyces quintiniae (incl. Linteromyces gen. nov.) on leaves of Quintinia sieberi, Lophotrichus medusoides from stem tissue of Citrus garrawayi, Mycena pulchra on soil, Neocalonectria tristaniopsidis (incl. Neocalonectria gen. nov.) and Xyladictyochaeta tristaniopsidis on leaves of Tristaniopsis collina, Parasarocladium tasmanniae on leaves of Tasmannia insipida, Phytophthora aquae-cooljarloo from pond water, Serendipita whamiae as endophyte from roots of Eriochilus cucullatus, Veloboletus limbatus (incl. Veloboletus gen. nov.) on soil. Austria, Cortinarius glaucoelotus on soil. Bulgaria, Suhomyces rilaensis from the gut of Bolitophagus interruptus found on a Polyporus sp. Canada, Cantharellus betularum among leaf litter of Betula, Penicillium saanichii from house dust. Chile, Circinella lampensis on soil, Exophiala embothrii from rhizosphere of Embothrium coccineum. China, Colletotrichum cycadis on leaves of Cycas revoluta. Croatia, Phialocephala melitaea on fallen branch of Pinus halepensis. Czech Republic, Geoglossum jirinae on soil, Pyrenochaetopsis rajhradensis from dead wood of Buxus sempervirens. Dominican Republic, Amanita domingensis on litter of deciduous wood, Melanoleuca dominicana on forest litter. France, Crinipellis nigrolamellata (Martinique) on leaves of Pisonia fragrans, Talaromyces pulveris from bore dust of Xestobium rufovillosum infesting floorboards. French Guiana, Hypoxylon hepaticolor on dead corticated branch. Great Britain, Inocybe ionolepis on soil. India, Cortinarius indopurpurascens among leaf litter of Quercus leucotrichophora. Iran, Pseudopyricularia javanii on infected leaves of Cyperus sp., Xenomonodictys iranica (incl. Xenomonodictys gen. nov.) on wood of Fagus orientalis. Italy, Penicillium vallebormidaense from compost. Namibia, Alternaria mirabibensis on plant litter, Curvularia moringae and Moringomyces phantasmae (incl. Moringomyces gen. nov.) on leaves and flowers of Moringa ovalifolia, Gobabebomyces vachelliae (incl. Gobabebomyces gen. nov.) on leaves of Vachellia erioloba, Preussia procaviae on dung of Procavia capensis. Pakistan, Russula shawarensis from soil on forest floor. Russia, Cyberlindnera dauci from Daucus carota. South Africa, Acremonium behniae on leaves of Behnia reticulata, Dothiora aloidendri and Hantamomyces aloidendri (incl. Hantamomyces gen. nov.) on leaves of Aloidendron dichotomum, Endoconidioma euphorbiae on leaves of Euphorbia mauritanica, Eucasphaeria proteae on leaves of Protea neriifolia, Exophiala mali from inner fruit tissue of Malus sp., Graminopassalora geissorhizae on leaves of Geissorhiza splendidissima, Neocamarosporium leipoldtiae on leaves of Leipoldtia schultzii, Neocladosporium osteospermi on leaf spots of Osteospermum moniliferum, Neometulocladosporiella seifertii on leaves of Combretum caffrum, Paramyrothecium pituitipietianum on stems of Grielum humifusum, Phytopythium paucipapillatum from roots of Vitis sp., Stemphylium carpobroti and Verrucocladosporium carpobroti on leaves of Carpobrotus quadrifolius, Suttonomyces cephalophylli on leaves of Cephalophyllum pilansii. Sweden, Coprinopsis rubra on cow dung, Elaphomyces nemoreus from deciduous woodlands. Spain, Polyscytalum pini-canariensis on needles of Pinus canariensis, Pseudosubramaniomyces septatus from stream sediment, Tuber lusitanicum on soil under Quercus suber. Thailand, Tolypocladium flavonigrum on Elaphomyces sp. USA, Chaetothyrina spondiadis on fruits of Spondias mombin, Gymnascella minnisii from bat guano, Juncomyces patwiniorum on culms of Juncus effusus, Moelleriella puertoricoensis on scale insect, Neodothiora populina (incl. Neodothiora gen. nov.) on stem cankers of Populus tremuloides, Pseudogymnoascus palmeri from cave sediment. Vietnam, Cyphellophora vietnamensis on leaf litter, Tylopilus subotsuensis on soil in montane evergreen broadleaf forest. Morphological and culture characteristics are supported by DNA barcodes.

Nemaline myopathy in a six-month-old Pomeranian dog.

ABSTRACT: Nemaline myopathy - a clinically and genetically complex heterogenous group of disorders - is described uncommonly in humans and rarely in animals, and is characterised by progressive muscle weakness. The diagnosis is confirmed by histological and/or ultrastructural identification of subsarcolemmal, thread-like, rod-shaped structures called nemaline rod bodies within more than 40% of skeletal muscle fibres. These rods contain the Z-line protein, α-actinin, that can be effectively stained in skeletal muscles using Gomori or Masson trichrome and negatively stained with periodic acid-Schiff. Similar rod-like bodies have been found in smaller numbers in dogs with endocrine disorders and occasionally in other conditions in humans. This report is of a six-monthold Pomeranian dog which had progressive exercise intolerance over a two-month period associated with severe disuse muscle atrophy of the thoracic limbs, as well as gradual pelvic limb weakness and regurgitation of food. Baseline diagnostics ruled out endocrinopathies and after histological and ultrastructural evaluation of thoracic limb muscles and nerve biopsies confirmed nemaline myopathy. The clinical course, diagnostic test results, ultrastructure of skeletal muscle and peripheral nerve, gross necropsy findings and histopathology using various stains are described and illustrated.

Fusarium graminearum Species Complex Associated with Maize Crowns and Roots in the KwaZulu-Natal Province of South Africa.

Thirty-three isolates of the Fusarium graminearum species complex obtained from diseased maize (Zea mays) crowns and roots in the Winterton district, KwaZulu-Natal province of South Africa were identified to species level. Their pathogenicity and virulence to maize 'PHI 32D96B' seedlings were determined under glasshouse conditions, with seedling survival and growth and crown and root rot as criteria. Phylogenetic analyses using the 3-O-acetyltransferase (Tri101) gene region sequences revealed the presence of F. boothii (2 isolates), F. graminearum sensu stricto (26 isolates), and F. meridionale (5 isolates) in the F. graminearum species complex associated with diseased maize crowns and roots. Pathogenicity results showed that F. boothii was the most and F. meridionale the least virulent of the three species. F. boothii and F. graminearum sensu stricto significantly reduced survival of seedlings and all three species caused significant reduction in growth and significantly more crown and root rot than the control (uninoculated). This is the first report of F. boothii, F. graminearum sensu stricto, and F. meridionale associated with diseased maize crowns and roots and their pathogenicity and virulence as soilborne pathogens on maize seedlings in South Africa.

Prevalence of subclinical bactibilia in apparently healthy shelter dogs.

OBJECTIVES: To determine the prevalence of subclinical bactibilia in apparently healthy shelter dogs and characterise serum liver enzymes activities, hepatobiliary histopathology and bile cytology in apparently healthy dogs with and without bactibilia. MATERIALS AND METHODS: Healthy, abandoned dogs euthanased for non-medical reasons were prospectively recruited for this cross-sectional study. Whole blood, collected immediately before euthanasia, was submitted for serum liver enzyme activity (alkaline phosphatase, alanine aminotransferase and gamma-glutamyl transferase) analyses. Bile, gall bladder and liver samples were collected aseptically from dogs within 25 minutes of euthanasia. Bile was submitted for bacterial culture and cytology in all dogs. Gall bladder and liver samples were submitted for histopathological examination in bactibilic dogs and nine randomly selected non-bactibilic dogs. RESULTS: Sixty-five healthy dogs were included in this study. Bactibilia was diagnosed in 10.8% (7/65) of dogs, with 9.2% (6/65) of dogs diagnosed on cytological examination and 4.6% (3/65) on culture. Elevated alanine aminotransferase activities were present in one bactibilic and five non-bactibilic dogs; and elevated gamma-glutamyl transferase activities in one bactibilic and two non-bactibilic dogs. No dogs had elevated alkaline phosphatase activities. Histopathological changes in bactibilic and non-bactibilic dogs included lymphoplasmocellular cholecystitis (7/7 and 9/9), gall bladder oedema (7/7 and 9/9), hepatic vacuolar degeneration (6/7 and 8/9), cholangitis (5/7 and 7/9), hepatic nodular hyperplasia (3/7 and 5/9) and hepatic cholestasis (2/7 and 4/9). CLINICAL SIGNIFICANCE: This study confirms that subclinical bactibilia occurs in a small number of apparently healthy shelter dogs and that subclinical hepatobiliary histopathological abnormalities can occur in apparently healthy bactibilic and non-bactibilic dogs.

Isolation and Identification of the Causal Agent of Brown Stalk Rot, A New Disease of Maize in South Africa.

During 2004 to 2005, an unreported disease of maize (Zea mays) was observed on commercial fields in the Northwest and Mpumalanga Provinces of South Africa. Infected plants were stunted, with a vertical crack at the first internode. Inside the stem, a dark-brown, narrow lesion was present along the crack. Internal browning inside the stem extended upward, reaching the top internode in some plants. Seed cobs were underdeveloped. Diseased plants were scattered in the fields and 10 to 70% of the crop was affected. Gram-negative, facultatively anaerobic bacteria were consistently isolated from diseased tissues. Pathogenicity tests established that representative strains induced disease symptoms similar to those observed on maize plants in the field. Physiological and biochemical characterization using the API 20E and API 50CHE systems and 16S rRNA gene sequence analyses showed that the strains belonged to the genus Pantoea. The results of these tests also separated the strains into two groups. The first group, giving a positive reaction in the indole test, was similar to Pantoea ananatis. The second group of strains was indole negative and resembled P. agglomerans. The fluorescent amplified fragment length polymorphism (F-AFLP) genomic fingerprints generated by the indole-positive strains and P. ananatis reference strains were similar and clustered together in the dendrogram, confirming that the indole-positive bacteria causing brown stalk rot on maize were P. ananatis. The F-AFLP fingerprints produced by the indole-negative strains were distinctly different from those generated by P. ananatis, P. agglomerans, P. dispersa, P. citrea, P. stewartii subsp. stewartii, and P. stewartii subsp. indologenes. The results indicated that indole-negative bacteria causing brown stalk rot on maize might belong to a previously undescribed species of the genus Pantoea. This is the first report of a new disease on maize, brown stalk rot, caused by two bacterial species, P. ananatis and an undescribed Pantoea sp.

Colony variation in Sinorhizobium meliloti inoculant strain U 45.

A culture of Sinorhizobium meliloti strain U 45, maintained on yeast extract-mannitol (YM) agar, produced a mixture of Congo red-absorbing (R1) and non-absorbing (W1) colonies when grown on YM medium containing Congo red. The original freeze-dried (FD) culture formed gummy (G), white (W2) and small red (R2) colony types on the above medium. All colonies were stable except G, which segregated into G and W2-like types. Immune diffusion patterns of all colony types were identical. The W1 colony type dominated R1 when a 1:1 combination was sub-cultured on YM agar. The parent cultures and their variants exhibited a range of N2-fixing effectiveness and competitiveness when inoculated onto two cultivars of Medicago sativa. Variant R2 from the FD culture was ineffective on both cultivars. Genomic DNA fingerprinting with insertion elements ISRm3 and ISRm2011-2 suggested that transposition of these elements was not a cause of variation, but a DNA band was absent in the profiles of two out of three W2-like colonies. Protein profile comparisons showed high similarity (r = 0.98) between the colony types when grown in YM broth. When grown on Tryptone-Yeast extract medium, variants from the FD and agar-maintained cultures formed separate clusters with r = 0.79. Polymerase chain reaction fingerprinting using repetitive, site-directed and arbitrary primers failed to differentiate the variants. The results emphasize the need to monitor culture variability to maintain the quality of legume inoculants.

First Report of Stem and Root Rot of Tomato Caused by Phytophthora capsici in South Africa.

Tomato plants, grown in open hydroponic systems under shadecloth and plastic near Barberton and Pretoria in South Africa and Srelebi Phikwe in Botswana, developed symptoms of wilting with brown-to-black cankers on the lower stems, blackening of the vascular tissues, and root rot. Pathogens isolated from affected tissues were identified as Phytophthora capsici Leonian (1) and Pythium aphanidermatum (Edson) Fitzp. (2).They occurred separately or together. Pythium aphanidermatum has previously been recorded on tomato in South Africa. P. capsici isolates were papillate, caducous, grew at >36°C, had tapered sporangial bases, and a maximum sporangial length of >70 µm. Koch's postulates were confirmed by inoculating 4-week-old tomato seedlings (cv. Floradade) grown at 22 to 30°C in a steam-pasteurized mixture of sawdust compost, pine bark, and vermiculite (3:2:1). Plugs from V8 juice agar cultures of P. capsici were placed on wounds made on the stems of 10 seedlings. Ten wounded uninoculated plants served as controls. Water-soaked lesions were visible on the stems of all inoculated plants after 2 days. Control plants remained healthy. After 4 days, lesions turned dark brown with affected plants wilted or dead. Reisolation yielded P. capsici. The experiment was repeated with similar results. To our knowledge, this is the first report of P. capsici on tomatoes in South Africa. References: (1) A. H. Thompson et al. S. Afr. J. Bot. 60:257, 1994.(2) W. Dick. Keys to Pythium. University of Reading Press, Reading, U.K., 1990.

First Report of Basal Stem Rot Caused by Pythium Group G on Kenaf in South Africa.

Kenaf (Hibiscus cannabinus L. Malvaceae) presents a source of high-quality cellulose fibers and is being investigated in South Africa for commercial production. In March 2001, 12 5-month-old kenaf plants grown from seed in experimental plots near Bloemfontein, South Africa, displayed large, black, sunken lesions (10 to 20 cm long) at the base of the stem, and severe root rot. A study was undertaken to characterize the pathogen, and to determine the relative susceptibilities of five kenaf genotypes being considered for commercial cultivation. Isolations from diseased tissue on malt extract agar consistently yielded a fungus identified as Pythium group G (1). Four-month-old kenaf plants were artificially inoculated in the field by inserting wooden toothpick tips colonized by the pathogen approximately 25 cm above soil level into the stems of 10 plants of each of five genotypes. Inoculation points were wrapped using Parafilm. The fungus was highly virulent to all five kenaf genotypes in two experiments, with mean cambial lesion lengths of 117, 119, 120, 122, and 139 mm at 7 days after inoculation for Tainung-2, Cuba 108, SF-459, El Salvador, and Everglades 41, respectively. Lesions ranged from 44 to 164 mm, with an overall mean of 124 mm for all five genotypes. No lesions developed in control plants. Although Everglades had the longest lesions, there were no significant differences (P < 0.05) among genotypes. Koch's postulates were completed by reisolating the fungus from all inoculated plants. To our knowledge, there are no published reports of Pythium group G causing stem or root rot of kenaf. References: (1) M. W. Dick. Keys to Pythium. University of Reading Press, Reading, UK, 1990.

Root Rot and Stunting of Hydroponically Grown Endive, Fennel, and Sorrel Caused by Pythium F-group in South Africa.

Specialized vegetable crops such as endive (Cichorium endiva), fennel (Foeniculum vulgare), and sorrel (Rumex spp.) are being cultivated hydroponically in South Africa to be marketed as admixtures in salads. Stunted growth accompanied by browning and rotting of the root tips has been observed at a commercial recirculating gravel bed hydroponic system near Pretoria during the warm summer months. Root segments excised from symptomatic plants were rinsed in sterile water and plated on Pythium selective medium (1). Pythium F-group, characterized by the production of noninflated filamentous sporangia and no oospores (2), was isolated from 40% of endive, 60% of fennel, and 7% of sorrel root segments. Koch's postulates were confirmed by inoculating 4-week-old seedlings of each crop in the greenhouse with a Pythium F-group isolate from the particular crop. Inoculations were performed by adding 3 ml of suspension (105 zoospores per ml) to each liter of aerated nutrient solution. Control plants received no inoculum, and the experiment was repeated once. After 4 weeks, inoculated plants showed stunting of the foliage and slight to moderate root rot. Pythium F-group could readily be reisolated from roots of inoculated plants. To our knowledge, this is the first report of Pythium F-group on these crops in South Africa. References: (1) W. J. Botha and R. L. J. Coetzer. S. Afr. J. Bot. 62:196, 1996. (2) M. W. Dick. Keys to Pythium. University of Reading Press, Reading, UK, 1990.

Pythium spp. Infecting Hydroponically Grown Lettuce in South Africa.

Root rot and wilting of lettuce (Lactuca sativa L. var. capitata L.) commonly occur in hydroponic systems in South Africa. Plants showing stunted growth, wilt, and root rot were collected at two commercial hydroponic systems in Gauteng Province, and root segments were plated on a Pythium selective medium (1). P. irregulare and three species groups without oogonia (F, HS, and T) were identified (2). Pythium groups F, HS, and T, and P. irregulare were represented by 63, 6, 5, and 26% of the isolates, repectively. Koch's postulates were confirmed in two greenhouse experiments by inoculating 4-week-old lettuce cv. Lutetia seedlings grown in aerated nutrient solution at 21°C. Two V8 juice agar cultures of an isolate of each species or species group were blended in 500 ml of sterile water, and 166 ml of suspension was added to the nutrient solution in each of three 5-liter receptacles containing four lettuce seedlings. Control plants received sterile V8 juice agar suspension. After 4 weeks, all Pythium isolates caused root rot and reduced fresh plant weight significantly, although no wilting occurred. Symptoms were more severe with P. irregulare and Pythium group HS than with groups F and T. Compared to the control, P. irregulare reduced fresh root and shoot weight of plants by 51 and 38%, Pythium group HS by 41 and 33%, Pythium group T by 29 and 26%, and Pythium group F by 30 and 24%, respectively. References: (1) W. J. Botha and R. L. J. Coetzer. S. Afr. J. Bot. 62:196, 1996. (2) M. W. Dick. Keys to Pythium. University of Reading Press, Reading UK, 1990.

Report of Root Rot Caused by Pythium F-Group on Hydroponically Grown Celery in South Africa.

Celery (Apium graveolens) is commonly grown in hydroponic systems in South Africa. During the warm summer months, plants often become stunted or die, with mortality as high as 70% in some systems. Affected plants show severe root rot and yellowing of the foliage. Root segments excised from symptomatic celery plants collected from various hydroponic systems in Gauteng Province, South Africa, were rinsed in sterile distilled water and plated on PARP medium (2). Pythium was isolated consistently from both rotted and symptomless roots. Isolates produced only noninflated filamentous sporangia and no oospores, conforming to the description of Pythium F-group (1). Koch's postulates were confirmed by inoculating 4-week-old celery seedlings cultivated in aerated nutrient solution in a hydroculture system in the greenhouse. A zoospore suspension (105 zoospores per ml) of Pythium F-group cultured on V8 juice agar was added to the nutrient solution at the rate of 3 ml/1iter. Control plants received no inoculum, and the experiment was repeated once. Plants were assessed after 4 weeks. All inoculated plants showed symptoms of root rot, stunting, and yellowing, whereas control plants remained healthy. Pythium F-group was recovered on PARP medium only from the roots of inoculated plants. Pythium F-group has been described as a pathogen of celery elsewhere, but to our knowledge, this is the first report of root rot caused by the fungus on celery in South Africa. References: (1) M. W. Dick. Keys to Pythium. University of Reading Press, Reading, UK, 1990. (2) S. N. Jeffers and S. B. Martin. Plant Dis. 70:1038, 1986.

Antibody response in cattle to oil emulsion rabies and ephemeral fever vaccines.

A stable oil emulsion rabies vaccine with a low viscosity was composed by a formula previously employed for Newcastle disease vaccine. Cattle developed high and sustained antibody levels, and guinea pigs were found to be solidly immune after a single injection of this vaccine. Antibody responses in cattle to 2 oil emulsion ephemeral fever vaccines were not satisfactory after a single injection, and severe local reactions were encountered when booster injections were applied.

The effect of passive immunization on active immunity against Clostridium perfringens type D in lambs.

Lambs in different stages of development of active immunity against Clostridium perfringens type D were treated with partially purified immunoglobulin in an attempt to superimpose a passive immunity on an existing or developing active immunity. Three different studies were undertaken to determine the impact of partial purified immunoglobulins on these vaccinated animals. In 2 of the 3 studies, active immunity was induced by administering the normal routine enterotoxaemia vaccinations and allowing the basic immunity to become established, for a period ranging from 2 weeks for the animals in study 1 and 4 months for those in study 2, before passive immunization with the partially purified immunoglobulins took place. An increase in the epsilon antibody titre occurred in each of the 2 studies after the animals were passively immunized with immunoglobulin, though this increase was not statistically significant (P greater than 0.05). In the 3rd study, when the animals were given the initial vaccination of the Onderstepoort enterotoxaemia oil adjuvant vaccine together with the immunoglobulin, an immediate increase in the epsilon antitoxin titre occurred that was statistically significant (P less than 0.05) 2-14 days after administration. No negative effects were noted on the development of an initial active immunity or an existing active immunity against Clostridium perfringens type D when they were passively immunized with partially purified immunoglobulin.

Studies of encephalitozoonosis in vervet monkeys (Cercopithecus pygerythrus) orally inoculated with spores of Encephalitozoon cuniculi isolated from dogs (Canis familiaris).

Encephlitozoonosis was induced in 35 of 38 vervet monkeys (Cercopithecus pygerythrus). They were either directly (orally) inoculated with Encephlitozoon cuniculi or indirectly exposed to this protozoan parasite. Cell-culture-grown spores of E. cuniculi, isolated from the kidneys of dogs with natural, fatal disease, were administered orally to 29 of these monkeys. Another 5 were exposed in utero by orally infecting pregnant females, and 3 were exposed to horizontal infection by nursing infected infants. Only one was given an intravenous inoculation of spores. The disease was induced in non-gravid and late-pregnant adults, immunocompetent infants, and in infants that were immunologically compromised by parenteral steroid administration, as well as in one infant that was immunologically immature because of its premature birth. The effects of age, dosage, post-inoculation (PI) interval, passage level of the parasite in cell culture and immunological status of the host were correlated with macroscopical and microscopical lesions. The experimentally induced infection was confirmed either by reisolation of the parasite in cell culture or by observation of spores in tissue sections. Both confirmatory methods were supported by serological examination. Reisolation of the organism in primary cell culture prepared from kidneys usually resulted in more frequent isolates and larger yields of spores from infants than from adult vervets. Infection with E. cuniculi invariably induced subclinical disease. Based on histology, lesions were minimal to moderately severe, depending on age, PI interval, and immunological status of the host. Alimentary tract infections were seen histologically as early as three days PI. Subsequently, infections resulted in detectable lesions most consistently in the liver, kidneys and brain. Lesions in these organs were generally granulomatous and were similar to those found in canine encephalitozoonosis. In addition, multifocal interstitial pneumonitis and myocarditis as well as vasculitis and perivasculitis were seen in other tissues and organs. Infants had more severe and more widespread lesions than adults. Although lesions and spores were still present in the brain of one immunocompetent infant 36 weeks after initial infection, the disease in immunocompetent infants and adults is thought to be self-limiting. However, infection may persist. Immunological depression favoured increased growth and multiplication of the organism, and resulted in detection of more spores within inflammatory lesions as well as more intracellular colonies of the organism that were free of inflammatory reaction.(ABSTRACT TRUNCATED AT 400 WORDS)

Composition and evaluation of the efficacy of Staphylococcus aureus vaccine.

An alum-precipitated Staphylococcus aureus vaccine, composed of a formalin-inactivated whole culture of a strain which produces Smith surface antigen and combined with the whole culture of a highly toxigenic strain, was found to afford a good immunity to staphylococcal skin infection in rabbits. Three injections of the vaccine provided immunity which lasted for at least 6 months against a primarily pyogenic strain of S. aureus and for at least 3 months against a toxigenic strain. From experiments using vaccines prepared from cells or toxoid only, it was deduced that, although there is a measure of strain specific immunity, a good heterologous immunity can be established with a combined product provided that it contains adequate quantities of toxoid. The use of such a vaccine as a potential aid in the control of bovine staphylococcal mastitis is discussed.

The passive protection of lambs against Clostridium perfringens type D with semi-purified hyperimmune serum.

Weaned lambs, having a detectable level of maternal antibodies (1-2 units/ml) against C. perfringens type D, showed protective antitoxin levels lasting for 29 days after receiving a single parenteral dose of 200 units/kg hyperimmune serum. Lambs, having no maternal antibodies (less than 0,07 units/ml) to C. perfringens type D but receiving the same dose of hyperimmune serum, maintained protective antibody levels for only 21 days. Three weeks after the titres fell below the minimum protective level of 0,15 units/ml, both these groups were treated again in the same manner. The passive immunity conferred in both groups now lasted for 42 days. When the hyperimmune serum was administered to lambs already immunized by vaccination, a slight increase was noted in the antibody titre.

Immunization of guinea-pigs and cattle with a reduced dose Clostridium chauvoei vaccine produced in a semi-synthetic medium.

A semi-synthetic culture medium and method are described for the production of a reduced dose Clostridium chauvoei vaccine. The vaccine gave excellent results in guinea-pigs, and 2 injections of 2.0 ml protected cattle against challenge with 2 M.L.D. of a virulent culture for at least 12 months. The suitability of C. chauvoei Strain OP64 as a vaccine strain was confirmed.

Antibody response to and immunity induced by Corynebacterium pyogenes vaccine.

A procedure is described for producing a high quality C. pyogenes vaccine. A satisfactory antibody response was obtained by administering 3 subcutaneous injections of vaccine at 10-day intervals, using doses of 2,0 ml for rabbits, 5,0 ml for sheep and 10,0 ml for cattle. The titres resulting from this procedure compared favourably with those obtained from following the more intensive schedule and higher doses of vaccine recommended by the British Veterinary Codex. The antibody response was at its best 1-2 months after immunization but dropped to a plateau 6 months after immunization. Mice were also successfully immunized against challenge with 1,5x108 organisms by the application of 3 doses of vaccine administered at 10-day intervals.

Comparison of oil adjuvant and aluminium phosphate-adsorbed toxoid for the passive immunization of lambs against tetanus.

Immunization of ewes with oil emulsion toxoid followed by an aluminium phosphate-adsorbed toxoid both containing 10 Lf per dose, resulted in a very high antitoxin level in their lambs. Two injections of aluminium phosphate-adsorbed toxoid also imparted a passive immunity to lambs which is considered to be adequate to protect them against tetanus for 4 weeks after birth.

Light and electron microscopical studies on canine encephalitozoonosis: cerebral vasculitis.

Brain lesions in 2 natural cases of canine encephalitozoonosis were studied by light and electron microscopy. Granulomatous reactions, associated with small vessels and capillaries, partly originated from extensive perithelial cell proliferation which ultimately produced the epithelioid cell component. Diffuse glial reactions apparently occurred in relation to the vasculitis. Lymphoid cells infiltrated the epithelioid and glial cell inflammation. Encephalitozoon in all its reproductive stages was identified as the aetiological agent, and ultrastructurally differentiated from Nosema on the basis that a single nucleus was observed. Viable organisms were present only within endothelial cells. Macrophages containing dead spores were usually seen around parasitized vessels and, less frequently, in the neuropil. Organisms, whether viable or non-viable, were never seen extracellularly at the ultrastructural level. Selected histochemical stains and electron microscopy were used to differentiate between viable and non-viable spores. Vasculitis is demonstrated as the underlying lesion of canine encephalitozoonosis affecting the brain and is suggested to be the basic factor in the pathogenesis of this disease.