Streptococcus dysgalactiae ssp. dysgalactiae in Norwegian bovine dairy herds: Risk factors, sources, and genomic diversity.

Despite the importance of Streptococcus dysgalactiae ssp. dysgalactiae (SDSD) as an udder pathogen, the reservoir and epidemiological characteristics of this bacterium are largely unexplored. The aims of this study were to investigate risk factors for SDSD intramammary infections (SDSD-IMI) in Norwegian bovine dairy herds, identify sources of SDSD on animals and in the environment, and elucidate the genetic diversity of SDSD isolates. Data from herd recordings and a questionnaire were used to investigate herd-level risk factors for SDSD-IMI in 359 freestall dairy herds. Seven herds with a suspected high prevalence of SDSD-IMI were visited to sample extramammary sources (e.g., skin, wounds, mucous membranes, and freestall environment). Bacterial isolates were whole-genome sequenced to investigate the distribution of SDSD genotypes within herds and to assess the phylogenetic relationship between SDSD isolates from 27 herds across Norway. Risk factors for high incidence of SDSD-IMI in freestall dairy herds were related to housing, including closed flooring in alleys and rubber mats in cubicle bases. Parlor milking was also a risk factor compared with automatic milking systems. From herd visits, a considerable proportion of extramammary samples were SDSD positive, particularly from wounds and skin of the animals and the cubicle bases. Samples from mucous surfaces (nostrils, rectum, and vagina) and water troughs were least frequently positive. Eight multilocus sequence types (ST) were identified among the sequenced isolates from 27 herds, and phylogenetic analyses revealed 8 clades corresponding to ST. No significant association was identified between sampling site (milk, body sites, and environment) and ST. In 4 of 6 herds from which 5 or more isolates were available, one ST dominated and was found in milk and extramammary samples. One ST (ST453) was found in 15 of 27 herds, which implies that this is a widely distributed and possibly a bovine-adapted strain. Findings in this study suggest that SDSD is a cow-adapted opportunist with potential for contagious transmission, and that the freestall environment is likely to play a role in transmission between cows.

Streptococcus agalactiae in the environment of bovine dairy herds--rewriting the textbooks?

Many free-stall bovine dairy herds in Norway fail to eradicate Streptococcus agalactiae despite long-term control measures. In a longitudinal study of 4 free-stall herds with automatic milking systems (AMS), milk and extramammary sites were sampled 4 times with 1-2 month intervals. Composite milk, rectal- and vaginal swabs were collected from dairy cows; rectal swabs from heifers and young stock; rectal- and tonsillar swabs from calves; and environmental swabs from the AMS, the floors, cow beds, watering and feeding equipment. A cross sectional study of 37 herds was also conducted, with 1 visit for environmental sampling. Fifteen of the herds were known to be infected with S. agalactiae while the remaining 22 had not had evidence of S. agalactiae mastitis in the preceding 2 years. All samples were cultured for S. agalactiae, and selected isolates (n=54) from positive herds were genotyped by Multi Locus Sequence Typing (MLST). Results show that the bovine gastrointestinal tract and the dairy cow environment are reservoirs of S. agalactiae, and point to the existence of 2 transmission cycles; a contagious transmission cycle via the milking machine and an oro-fecal transmission cycle, with drinking water as the most likely vehicle for transmission. Ten sequence types were identified, and results suggest that strains differ in their ability to survive in the environment and transmit within dairy herds. Measures to eradicate S. agalactiae from bovine dairy herds should take into account the extra-mammary reservoirs and the potential for environmental transmission of this supposedly exclusively contagious pathogen.

Acute and Chronic Erysipelothrix rhusiopathiae Infection in Lambs.

Polyarthritis caused by Erysipelothrix rhusiopathiae is a relatively common infection in lambs characterized by low mortality and high morbidity. E. rhusiopathiae is a ubiquitous Gram-positive bacterium that is both a commensal and a pathogen of vertebrates. The disease was studied during an outbreak in a Norwegian Spæl sheep flock. In the acute phase, 48 of 230 (20%) lambs developed clinical signs and 4 died (1.7%). One acute case was necropsied and E. rhusiopathiae was cultured from all major organs investigated and from joints. There was a fibrinous polyarthritis, increased presence of monocytes in vessels, and necrosis of Purkinje cells. Sixteen of the diseased animals (33%) developed a chronic polyarthritis. Eight of these lambs were necropsied; all had lesions in major limb joints, and 3 of 8 also had lesions in the atlanto-occipital joint. At this stage, E. rhusiopathiae was cultured only from the joints in 7 of 8 (87.5%) lambs, but by real-time polymerase chain reaction, we showed persistence of the bacterium in several organs. Pulsed-field gel electrophoresis typing of the bacterial isolates indicated that the same strain caused the acute and chronic disease. Five of 6 (83%) chronically affected animals had amyloidosis of the spleen, and 6 of 8 (75%) had amyloidosis of the liver. All chronically affected animals had a glomerulonephritis, and 6 of 8 (75%) had sparse degeneration in the brain. Ceruloplasmin and haptoglobin were significantly increased in the chronically diseased lambs. These results show that chronic ovine erysipelas is not restricted to joints but is a multisystemic disease.

Persistence of staphylococcal species and genotypes in the bovine udder.

Staphylococci are a major cause of intramammary infections (IMI) in ruminants. The main aim of this study was to investigate staphylococcal IMI in dairy cattle with emphasis on persistence and distribution of staphylococcal species and genotypes. With a sampling interval of 4-8 weeks, over a year, 4030 samples from 206 cows in 4 herds were collected. Coagulase-negative staphylococci (CNS) and Staphylococcus aureus were detected in 13.2% and 4.2% of the samples, respectively. Selected CNS isolates from quarter milk samples were identified to species level using sodA sequencing. Staphylococcus chromogenes (32%) and Staphylococcus simulans (25%) predominated. The proportion of S. chromogenes was greater in primiparous (52%) than in multiparous cows (12%), while the opposite was the case for Staphylococcus epidermidis (6% and 21%, respectively). Isolates from possibly persistent IMI were selected for pulsed-field gel electrophoresis (PFGE). Six staphylococcal species were found to cause persistent IMI; S. aureus, S. chromogenes, S. simulans, S. epidermidis, Staphylococcus haemolyticus and Staphylococcus warneri. It was shown that several pulsotypes (PTs) within each species were associated with persistent infections, but only a few were spread and caused persistent IMI in multiple cows within a herd. Of special interest was the observation that only one, or a few, strains of each species caused persistent IMI in multiple cows within a same herd. This indicates strain differences with respect to transmissibility and pathogenicity.

Reservoirs of Staphylococcus aureus in meat sheep and dairy cattle.

The objective of the study was to investigate reservoirs and transmission of S. aureus in ewes and lambs in 3 meat sheep flocks. Repeated sampling of milk, teat skin, nasal- and vaginal mucous membranes was performed and samples were analysed for S. aureus. For comparison, samples were also collected from cows and young heifers in 3 dairy cattle herds. Selected isolates were compared by pulsed-field gel electrophoresis (PFGE). S. aureus was detected in 8 (1.5%) of 520 milk samples from ewes and in 38 (6.4%) of 588 milk samples from cows. From body site swabs, S. aureus was found in 394 (32.6%) of 1208 samples from sheep and in 67 (16.0%) of 420 samples from cattle. The proportion of S. aureus-positive nasal swabs from ewes and cows were 56.7% and 13.9%, respectively. From lambs, 58.2% of the nasal swabs were S. aureus-positive. In each flock, one S. aureus pulsotype predominated. Identical S. aureus pulsotypes were found in milk and from body sites. Paired S. aureus isolates from the nasal cavity of (i) ewes and their lambs, (ii) twins and (iii) from repeated swabs of individual ewes were compared by PFGE, and in the majority of cases the two isolates were identical. The results contribute new knowledge indicating frequent transmission of S. aureus between the dam and her lambs and within animals in a flock. In contrast to cattle, S. aureus is frequently present in the nose of sheep which may represent the primary reservoir of S. aureus in sheep flocks.

First Report of Golovinomyces cichoracearum as the Causal Agent of Powdery Mildew on Symphyotrichum novi-belgii (Synonym Aster novi-belgii) in Denmark.

Symphyotrichum novi-belgii (L.) G.L. Nesom (synonym Aster novi-belgii L.) is an autumn flowering perennial used in gardens and as a cut flower. During the last 20 years, it has been developed as a potted plant, thereby increasing its economic importance. In Denmark, 7 to 8 million S. novi-belgii plants are produced annually, making it one of the 10 most popular potted plant crops ( http://floradania.dk/index.php?id=165 ). In general, S. novi-belgii is a healthy plant, but it can be severely attacked by powdery mildew both in greenhouse production and outdoors, and diseased plants have been observed in most parts of the country. Infected plants show typical symptoms: leaf surfaces become covered with white mycelium and as the disease progresses infected leaves turn yellow and die. Powdery mildew is regarded the main disease problem in S. novi-belgii and it causes problems year round in greenhouse production. Normally, the disease is controlled by fungicides, but once out of the production system, symptom development in the retail trade will reduce the plant's appeal to customers to a degree that prevents sales. The powdery mildew identified in this study was collected in a small research field at Aarslev, Denmark in September 2004. Since collection, the pathogen has been maintained in a greenhouse on S. novi-belgii and it has been used for disease resistance screening. However, lack of proper identification of the causal agent has hindered the development of powdery mildew resistant cultivars. To identify the pathogen, the internal transcribed spacer region (ITS) of the rDNA was amplified using primers ITS1 and ITS4 (2) and sequenced. The resulting sequence was deposited in GenBank (Accession No. HM769725). BLASTn analysis of the 598-bp fragment showed 99% identity to Golovinomyces cichoracearum (DC.) V.P. Heluta from Rudbeckia laciniata L. (Accession No. AB077622). The powdery mildew colonies were slightly pink with barrel-shaped, hyaline conidia borne in chains of three to four. The length of the conidia was 30 ± 4 µm and the width was 13 ± 1 µm (n = 105). Foot cells of the conidiophores were 101 ± 16 µm long and 12 ± 5 µm wide (n = 50) with a slight constriction at the base. Chasmothecia were not observed. These morphological characteristics confirmed the identification as G. cichoracearum (1). To fulfill Koch's postulates, 10 healthy S. novi-belgii 'Victoria Fanny' plants were inoculated in an inoculation tower by shaking infected S. novi-belgii plants over the tower, resulting in a spore density of 47 spores/mm2 on the leaf surface. The infected plants were placed in a growth chamber with 16 h of light (200 µmol·m-2·s-1) and day and night temperatures of 20 and 15°C, respectively. Symptoms developed on all plants after 11 days. Colony morphology on the leaves and the morphological characteristics were as described above. Conidia were washed off the leaves, DNA extracted, and the ITS was amplified by PCR. The resulting PCR product was sequenced and was identical to HM769725. To our knowledge, this is the first report of G. cichoracearum on S. novi-belgii in Denmark. References: (1) U. Braun. The Powdery Mildews (Erysiphales) of Europe. Gustav Fischer Verlag, Jena Germany, 1995. (2) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, New York, 1990.

Rhizobacterially induced protection of watermelon against Didymella bryoniae.

AIMS: To identify rhizobacteria from the Mekong Delta of Vietnam, which can systemically protect watermelon against Didymella bryoniae and elucidate the mechanisms involved in the protection conferred by isolate Pseudomonas aeruginosa 23(1-1). METHODS AND RESULTS: Bacteria were isolated from watermelon roots and their antagonistic ability tested in vitro. Of 190 strains, 68 were able to inhibit D. bryoniae by production of antibiotics. Four strains were able to reduce foliar infection by D. bryoniae when applied to watermelon seeds before sowing. Strain Ps. aeruginosa 23(1-1) was chosen for investigations of the mechanisms involved in protection and ability to control disease under field conditions. In the field, the bacterium was able to significantly reduce disease in two consecutive seasons and increase yield. Furthermore, it colonized watermelon plants endophytically, with higher numbers in plants infected by D. bryoniae than in noninoculated plants. To elucidate the mechanisms involved in protection, the infection biology of the pathogen was studied in bacterially treated and control plants. Pseudomonas aeruginosa 23(1-1) treatment inhibited pathogen penetration and this was associated with hydrogen peroxide accumulation, increased peroxidase activity and occurrence of new peroxidase isoforms, thus indicating that resistance was induced. CONCLUSIONS: The endophytic bacterium Ps. aeruginosa 23(1-1) can control D. bryoniae in watermelon by antibiosis and induced resistance under greenhouse and field conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings suggest that rhizobacteria from native soils in Vietnam can be used to control gummy stem blight of watermelon through various mechanisms including induction of resistance.

Bacteriological and molecular investigations of Staphylococcus aureus in dairy goats.

In order to investigate reservoirs of Staphylococcus aureus in dairy goats, samples for bacteriological analyses were collected from seven herds. S. aureus was detected in 353 (6.2%) of 5671 milk samples, 53 (9.9%) of 535 teat skin swabs, 392 (68.9%) of 569 nasal swabs and in 180 (31.6%) of 569 vaginal swabs. Vaginal swabs were more often S. aureus-positive after kidding (44.9%) than before drying off (19.1%), while nasal swabs were more often positive before drying off (75.6%) than after kidding (62.0%). Retrieved S. aureus isolates were compared by pulsed-field gel electrophoresis (PFGE), and selected isolates were tested for enterotoxin genes (se) by PCR. By PFGE, 505 S. aureus isolates were divided into 33 pulsotypes (PTs). The five most prevalent PTs included 73.3% of the isolates and were found in 3-5 herds. Pairs of S. aureus isolates from persistent intramammary infections (IMI), repeated vaginal swabs, and from milk and teat skin from the same animal were usually identical. Paired isolates from other body sites of the same animal, including from bilateral IMI, were identical in less than 50% of the situations. The majority (71.9%) of analysed S. aureus isolates were se-positive. The genes sec, sell and tst were detected almost exclusively, but no correlation was observed between persistence of IMI and the enterotoxin gene profile of the causal S. aureus strains. The frequent presence of S. aureus on the mucous membranes may contribute to dispersal of the bacteria among dairy goats, hampering effective transmission control in dairy goat herds.

Infection biology and defence responses in sorghum against Colletotrichum sublineolum.

AIMS: To investigate the infection biology of Colletotrichum sublineolum (isolate CP2126) and defence responses in leaves of resistant (SC146), intermediately resistant (SC326) and susceptible (BTx623) sorghum genotypes. METHODS AND RESULTS: Infection biology and defence responses were studied quantitatively by light microscopy, H(2)O(2) accumulation by DAB staining and HRGP accumulation by immunological methods. Inhibition of conidial germination and appressorium formation may represent prepenetration defence responses on the leaf surface. Inducible defence responses in the resistant genotypes included decreases in formation of appressoria as well as accumulation of H(2)O(2), HRGPs and phytoalexins. Concomitant with these inducible responses, fungal growth was stopped during or just after penetration in genotypes SC146 and SC326. High levels of H(2)O(2) accumulating at late infection stages (5 days after inoculation) in the susceptible genotype BTx623 correlated with necrosis and tissue degeneration. CONCLUSIONS: The early accumulation of H(2)O(2) and HRGPs indicates roles in defence whereas the late accumulation in genotype BTx623 correlated with successful pathogenesis. SIGNIFICANCE AND IMPACT OF THE STUDY: The fact that there is a significant correlation between induced accumulation of H(2)O(2), papilla formation and cell wall cross-linking, as evidenced by HRGP accumulation, and cessation of pathogen growth in resistant genotypes may help exploit host resistance in sorghum.

Genetic variation among Staphylococcus aureus strains from Norwegian bulk milk.

Strains of Staphylococcus aureus obtained from bovine (n = 117) and caprine (n = 114) bulk milk were characterized and compared with S. aureus strains from raw-milk products (n = 27), bovine mastitis specimens (n = 9), and human blood cultures (n = 39). All isolates were typed by pulsed-field gel electrophoresis (PFGE). In addition, subsets of isolates were characterized using multilocus sequence typing (MLST), multiplex PCR (m-PCR) for genes encoding nine of the staphylococcal enterotoxins (SE), and the cloverleaf method for penicillin resistance. A variety of genotypes were observed, and greater genetic diversity was found among bovine than caprine bulk milk isolates. Certain genotypes, with a wide geographic distribution, were common to bovine and caprine bulk milk and may represent ruminant-specialized S. aureus. Isolates with genotypes indistinguishable from those of strains from ruminant mastitis were frequently found in bulk milk, and strains with genotypes indistinguishable from those from bulk milk were observed in raw-milk products. This indicates that S. aureus from infected udders may contaminate bulk milk and, subsequently, raw-milk products. Human blood culture isolates were diverse and differed from isolates from other sources. Genotyping by PFGE, MLST, and m-PCR for SE genes largely corresponded. In general, isolates with indistinguishable PFGE banding patterns had the same SE gene profile and isolates with identical SE gene profiles were placed together in PFGE clusters. Phylogenetic analyses agreed with the division of MLST sequence types into clonal complexes, and isolates within the same clonal complex had the same SE gene profile. Furthermore, isolates within PFGE clusters generally belonged to the same clonal complex.

The occurrence of Staphylococcus aureus on a farm with small-scale production of raw milk cheese.

In recent years, the small-scale production of raw milk products has increased in Norway, and there is some concern that such foods may pose a risk of staphylococcal food poisoning to consumers. The aim of the study was to evaluate potential sources of contamination of raw milk cheese with Staphylococcus aureus on a bovine dairy farm with small-scale production. Samples for bacteriological analyses (n = 144) were collected from the animals, the environment, processing equipments, from humans, and from cheeses at various stages of production. Staphylococcus aureus was isolated from 10 of 11 cows, the farmer, equipment, the environment, and the cheese. Seventy-five Staph. aureus isolates were genotyped by pulsed-field gel electrophoresis, tested for enterotoxin (SE) production by reversed passive latex agglutination, for SE genes by multiplex polymerase chain reaction, and for penicillin resistance by the cloverleaf method. Five different pulsotypes were identified and SE gene fragments were identified in 11 isolates, but no isolates produced SE or were penicillin resistant. Staphylococcus aureus was found throughout the farm, and appeared to be spread with the milk to the environment, equipment, and to products. One pulsotype dominated and was identified from most sample sites on the farm. Raw milk products are vulnerable to contamination with Staph. aureus. Strategies to reduce the occurrence of Staph. aureus in bulk milk are of particular importance on farms where milk is used for raw milk products.

Comparison of Staphylococcus aureus genotypes recovered from cases of bovine, ovine, and caprine mastitis.

Staphylococcus aureus is an important pathogen in domestic ruminants. The main objective of this study was to determine the similarity of epidemiologically unrelated S. aureus isolates from bovine, ovine, and caprine mastitis. By pulsed-field gel electrophoresis, 160 different pulsotypes (PTs) were identified among 905 isolates recovered from 588 herds in 12 counties in Norway. Based on estimates of similarity, using an 80% cluster cutoff, the isolates were assigned to 47 clusters. One cluster included 62% of all the isolates and more than 45% of the isolates from each host species. Twenty-three PTs included isolates from more than one host species; these 23 PTs represented 72% of all the isolates. The six most prevalent PTs included isolates from all host species and contained 45% of the bovine isolates, 54% of the ovine isolates, and 37% of the caprine isolates. Antimicrobial susceptibility testing of 373 of the isolates revealed resistance to penicillin in 2.9% and to streptomycin in 2.4%; only 1.9% were resistant to 1 of the other 11 antimicrobials tested. The results of this study suggest that a small number of closely related genotypes are responsible for a great proportion of S. aureus mastitis cases in cows, ewes, and goats in Norway and that these genotypes exhibit little or no host preference among these species. Selection due to antimicrobial resistance appears not to have contributed to the predominance of these genotypes.

Enterotoxigenic Staphylococcus aureus in bulk milk in Norway.

AIMS: To investigate the presence of enterotoxigenic Staphylococcus aureus in bulk milk and in a selection of raw milk products. METHODS AND RESULTS: Samples of bovine (n = 220) and caprine (n = 213) bulk milk, and raw milk products (n = 82) were analysed for S. aureus. Isolates were tested for staphylococcal enterotoxin (SE) production (SEA-SED) by reversed passive latex agglutination and for SE genes (sea-see, seg-sej) by multiplex PCR. Staphylococcus aureus was detected in 165 (75%) bovine and 205 (96.2%) caprine bulk milk samples and in 31 (37.8%) raw milk product samples. Enterotoxin production was observed in 22.1% and 57.3% of S. aureus isolates from bovine and caprine bulk milk, respectively, while SE genes were detected in 52.5% of the bovine and 55.8% of the caprine bulk milk isolates. SEC and sec were most commonly detected. A greater diversity of SE genes were observed in bovine vs caprine isolates. CONCLUSIONS: Staphylococcus aureus seems highly prevalent in Norwegian bulk milk and isolates frequently produce SEs and contain SE genes. Enterotoxigenic S. aureus were also found in raw milk products. SIGNIFICANCE AND IMPACT OF THE STUDY: Staphylococcus aureus in Norwegian bovine and caprine bulk milk may constitute a risk with respect to staphylococcal food poisoning from raw milk products.

Diversity of Staphylococcus aureus enterotoxin types within single samples of raw milk and raw milk products.

AIM: To find out if testing of up to 10 Staphylococcus aureus isolates from each sample from raw milk and raw milk products for staphylococcal enterotoxin (SE) might increase the chances of identifying potential sources of food intoxication. METHODS AND RESULTS: Altogether 386 S. aureus isolates were tested for the presence of SE by reversed passive latex agglutination (SET-RPLA), and SE genes (se) by a multiplex polymerase chain reaction (PCR). In 18 of 34 (53%) S. aureus positive samples a mixture of SE and/or se positive and negative isolates were identified. Multiplex PCR increased the number of potential SE producing strains, i.e. isolates that harboured se, with 51% among the product and 48% among the raw bovine milk isolates. Examination by pulsed-field gel electrophoresis mostly confirmed clonal similarity among isolates sharing SE/se profile, but did not further differentiate between them. CONCLUSIONS: Isolates of S. aureus collected from one sample may show great diversity in SE production and different plating media seem to suppress or favour different strains of S. aureus. SIGNIFICANCE AND IMPACT OF THE STUDY: Several isolates of S. aureus from each sample should be tested for enterotoxin production in cases with typical SE intoxication symptoms with methods that are able to reveal new SE/se.

Mechanisms of Induced Resistance in Barley Against Drechslera teres.

ABSTRACT Quantitative and qualitative histopathological methods and molecular analyses were used to study the mechanisms by which preinoculation with either of the nonbarley pathogens, Bipolaris maydis and Septoria nodorum, inhibited growth of Drechslera teres. Collectively, our data suggest that induced resistance is the principal mechanism responsible for impeding the pathogen. The enhancement of resistance in the host was primarily manifested during penetration by D. teres, and after penetration, where growth of D. teres ceased soon after development of infection vesicles. Thus, 24 h after pretreatment with B. maydis or S. nodorum, the penetration frequency from D. teres appressoria was reduced from 42.7% in the controls to 9.5 and 14.8%, respectively. The reductions were associated with increased formation of fluorescent papillae in induced cells (early defense reaction). The postpenetrational inhibition of D. teres completely stopped fungal growth and was apparently linked to an enhancement of multicellular hypersensitive responses in inducer-treated leaves (late defense reaction). Papillae formation and multicellular hypersensitive reactions were also observed in fully susceptible, noninduced control leaves, but they were inadequate to stop fungal progress. Northern blots from leaves treated with either inducer alone support the conclusion that induced resistance is involved in suppression of D. teres by increased formation of papillae and hypersensitive reactions. Thus, the blots showed strong expression of several defense response genes that are involved in these reactions in barley attacked by Erysiphe graminis f. sp. hordei.

Suppression of Rice Blast by Preinoculation with Avirulent Pyricularia oryzae and the Nonrice Pathogen Bipolaris sorokiniana.

ABSTRACT Avirulent isolates of Pyricularia oryzae and isolates of Bipolaris sorokiniana, a nonrice pathogen, were used to suppress rice blast caused by P. oryzae. In greenhouse experiments, both fungi substantially reduced leaf blast when applied 24 h or more before the pathogen. B. sorokiniana, but not avirulent isolates of P. oryzae, systemically reduced disease in leaf 5 when applied to whole plants at the four-leaf stage. In field experiments, both fungi were able to reduce neck blast significantly. No increase in grain yield was obtained by using avirulent isolates of P. oryzae, whereas five sprays with B. sorokiniana from seedling to heading stages increased the grain yield in two of three experiments conducted at two locations in Nepal. The significant increase in yield was observed under high inoculum pressure of P. oryzae. Induced resistance is suggested to be involved in the suppression of disease.

Seedborne Infection of Rice by Pyricularia oryzae and Its Transmission to Seedlings.

Seedborne infection of rice by Pyricularia oryzae and its transmission to seedlings were studied quantitatively with naturally infected seeds of three rice cultivars collected from three locations in Nepal. A linear relationship on a logistic scale was found between panicle symptoms and seed infection, i.e., the more symptoms the higher seed infection. However, healthy-looking panicles and branches of panicles could also yield infected seeds. Postharvest measures such as winnowing and sun-drying significantly reduced seed infection by P. oryzae and filled grains had a lower degree of infection than unfilled grains. Sporulation of P. oryzae was most often confined to the embryonal end of germinating seeds. In contrast, most of the nongerminating seeds had sporulation all over the seed surface. Transmission of P. oryzae from seeds to seedlings, studied under various seeding conditions, showed that the transmission rate was always low. Thus, a seed sample with 21% seed infection resulted in less than 4% seedlings with blast lesions. Seed transmission was found for light covering of the seeds with soil or for moist seeding without covering. Transmission was rarely found when seeds were completely covered, and never in seedlings raised under water seeding conditions. Lower infection frequency was observed in seedlings raised in unsterilized soil than in seedlings raised in sterilized soil. Also, percent recovery of P. oryzae from infected seeds was higher in sterilized soil than in unsterilized soil and declined with time. Seedlings grown under low temperature (15 to 20°C) conditions did not develop blast lesions but when the same plants were transferred to high temperature (25 to 30°C) conditions, blast lesions were detected. This confirmed the latent infection in seedlings by P. oryzae grown under low temperature conditions.