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Each year, bovine respiratory disease (BRD) results in significant economic loss in the cattle sector, and novel metabolic profiling for early diagnosis represents a promising tool for developing effective measures for disease management. Here, 1 H-nuclear magnetic resonance ( 1 H-NMR) spectra were used to characterize metabolites from blood plasma collected from male dairy calves (n = 10) intentionally infected with two of the main BRD causal agents, bovine respiratory syncytial virus (BRSV) and Mannheimia haemolytica (MH), to generate a well-defined metabolomic profile under controlled conditions. In response to infection, 46 metabolites (BRSV = 32, MH = 33) changed in concentration compared to the uninfected state. Fuel substrates and products exhibited a particularly strong effect, reflecting imbalances that occur during the immune response. Furthermore, 1 H-NMR spectra from samples from the uninfected and infected stages were discriminated with an accuracy, sensitivity, and specificity ≥ 95% using chemometrics to model the changes associated with disease, suggesting that metabolic profiles can be used for further development, understanding, and validation of novel diagnostic tools., (© 2023. The Author(s).)

Powdery mildews (PMs) are important plant pathogens causing widespread damage. Here, we report the first draft genome of Erysiphe pulchra , the causative agent of PM of flowering dogwood, Cornus florida . The assembled genome was 63.5 Mbp and resulted in formation of 19,442 contigs (N50 = 11,686 bp) that contained an estimated 6,860 genes with a genome coverage of 62×. We found 102 candidate secreted effector proteins (CSEPs) in E. pulchra similar to E. necator genes that are potentially involved in disease development. This draft genome is an initial step for understanding the evolutionary history of the PMs and will also provide insight into evolutionary strategies that led to the wide host expansion and environmental adaptations so effectively employed by the PM lineages.

Twelve simple sequence repeat (SSRs) loci were used to evaluate genetic diversity of 109 isolates of Macrophomina phaseolina collected from different geographical regions and host species throughout the United States (US). Genetic diversity was assessed using Nei's minimum genetic distance, and the usefulness of each locus was determined by calculating the polymorphism information content (PIC). A total of 98 alleles were detected and of these 31 were unique to individual genotypes. Eight of twelve loci were highly informative with PIC values greater than 0.50. The majority of pairwise comparisons of genetic distance were greater than 0.60 indicating moderate to high genetic diversity. Dendrograms based on the genetic dissimilarities were created for the 109 isolates of which 79 were from soybean. Some clustering by host and geography was noted, but, the dendrograms generally grouped isolates independent of host or geography. Additionally, sequencing of the internal transcribed spacer region (ITS) for 10 isolates revealed that all of these isolates were 99% similar. Three SSR loci from M. phaseolina were cross amplified in other genera in the Botryosphaeriaceae. This was the first study of genotyping and assessing genetic diversity of M. phaseolina isolates collected from a widespread host and geographic range across the US with SSRs. With an additional 34 loci publically available for M. phaseolina, the results indicate that previously developed SSRs from one species can be used in future population, ecological, and genetic studies of M. phaseolina and other genera within the Botryosphaeriaceae.

Microsatellite loci were identified from the charcoal rot fungus (Macrophomina phaseolina). Primer pairs for 46 loci were developed, and of these, 13 were optimized and screened using genomic DNA from 55 fungal isolates collected predominantly from two soybean fields in Mississippi. Twelve of the optimized loci were polymorphic and the number of alleles per locus ranged from 6 to 22. These microsatellites will be useful in population and pathogenicity studies to correspond with development of potential disease-resistant soybean and other susceptible crops., (© 2009 Blackwell Publishing Ltd. No claim to original US government works.)

Loblolly pine (Pinus taeda) is the major commercial pine species cultivated in the Gulf Coast Region of the southern United States. Symptoms of Diplodia shoot blight (including yellow and brown needles and resin-soaked, dead, small twigs), pycnidia with conidia typical of Diplodia pinea on blighted shoots, and damaged, immature seed cones were observed during the summer of 2007 in loblolly pine seed orchards near Ward, AL, Winn Parish, LA, and Moselle, MS. Similar conidia also were obtained from pycnidia on opened seed cones of longleaf pine (P. palustris) collected on the campus of Mississipi State University, Starkville. Pure cultures obtained from specimens collected at each location were confirmed as D. pinea using species-specific PCR primers (3) that allow differentiation of D. pinea from the similar pine shoot blight pathogen D. scrobiculata. Isolates from loblolly pines in Alabama (07-58), Louisiana (07-38), and Mississippi (06-45) were used individually to inoculate potted 6- to 7-month-old loblolly pine seedlings grown from seed in a greenhouse in each of two independent trials. Elongating terminal shoots of seedlings to be inoculated were wounded by removing a needle fascicle ∼2 cm below the shoot apex. A 4-mm-diameter plug cut from an actively growing colony on water agar (WA) was placed on the wound, mycelium side toward the stem. Noncolonized WA plugs were placed in the same manner on similarly wounded control seedlings and nonwounded control seedlings also were used. Parafilm was wrapped around the shoots to hold the agar plugs in place and was removed after 1 week. Each of the five isolate-treatment combinations was applied to seven (trial 1) or eight (trial 2) seedlings (35 and 40 seedlings per trial, respectively). One week after inoculation, small, brown lesions were visible at the point of inoculation on stems of most of the inoculated seedlings. At 25 days after inoculation, all inoculated seedlings exhibited needle browning and stem cankers ranging from 0.6 cm to 9.0 cm long (mean 2.5 cm) that girdled and killed distal portions of the shoots of ∼25% of the inoculated seedlings in each trial. Wounded control and nonwounded control seedlings did not develop symptoms. Stem segments including the point of inoculation (or comparable segments of wounded and nonwounded control seedlings) were excised, surface disinfested, and incubated on tannic acid agar with sterile red pine needles. D. pinea was cultured from all inoculated seedlings and also from one wounded control seedling. Although occurrence of D. pinea on Cedrus spp. is included in an index (1), to our knowledge this is the first confirmed report of D. pinea on pines in Alabama, Louisiana, and Mississippi. The degree of risk presented by D. pinea to loblolly pine, longleaf pine, and other pine species native to the southern United States when grown in their native ranges is unknown. Reports of Diplodia shoot blight of southern U.S. pines when grown as exotics in the southern hemisphere (4) and the potential for epidemics to develop suddenly under severe weather conditions (2,4) justify additional studies to evaluate the potential for damage to these hosts in their native ranges. References: (1) Anonymous. Page 333 in: Index of Plant Diseases in the United States. Agric. Handb. 165, U.S. Dep. Agric. Washington, DC, 1960. (2) T. H. Nicholls and M. E. Ostry. Plant Dis. 74:54, 1990. (3) D. R. Smith and G. R. Stanosz. Plant Dis. 90:307, 2006. (4) W. J. Swart and M. J. Wingfield. Plant Dis. 75:761, 1991.

Research on charcoal rot disease in soybeans, and approximately 500 other plant diseases caused by the fungus Macrophomina phaseolina, has been severely hampered by unavailability of phaseolinone (1), an eremophilane sesquiterpenoid phytotoxin proposed to facilitate initial infection. Phytotoxin produced in cultures of disease-causing M. phaseolina isolated in Mississippi, and purified in a manner similar to that reported for 1, was shown to be (-)-botryodiplodin (2), a readily synthesized mycotoxin previously isolated from Botryodiplodia theobromae cultures. Phaseolinone was not detected, suggesting that 2 may be the phytotoxin that facilitates infection. The availability of 2 should facilitate studies on its role in plant disease.

Aspergillus flavus is a filamentous fungus that produces mycotoxins in many food and feed crops, such as maize (Zea mays L.). Isolates were analyzed for toxin production by nucleic acid profiles in an attempt to differentiate aflatoxigenic from nonaflatoxigenic isolates. A total of 41 aflatoxigenic and 34 nonalfatoxigenic isolates were included in the study. The isolates were evaluated initially using DNA amplification fingerprinting (DAF) without clear resolution of the groups. A weak association of aflatoxigenic isolates was observed, as evidenced by their clustering in 18 of 81 trees recovered from maximum parsimony analysis of binary characters derived from arbitrary signatures from amplification profiles (ASAP) data; nonaflatoxigenic isolates exhibited a pattern of paraphyletic laddering. Up to five markers unambiguously supported the aflatoxigenic isolate grouping, but the presence of alternative conflicting topologies in equally parsimonious trees precluded the observation of meaningful statistical support. With additional markers for genome of A. flavus, this method could be used to resolve toxigenic from nontoxigenic strains. This additional work could resolve aflatoxigenic isolates of A. flavus present on maize plants using ASAP, which would reduce labor intense costs and potentially lead to faster determination of resistant cultivars in breeding efforts.

Baseline information on the diversity and population densities of fungi collected from soil debris and cotton (Gossypium hirsutum L.) roots was determined. Samples were collected from Tifton, GA, and Starkville, MS containing cotton field soil treated with the nematicides 1,3-dichloroproprene (fumigant) and aldicarb (granules). A total of 10,550 and 13,450 fungal isolates were collected from these two study sites, respectively. Of this total, 34 genera of plant pathogenic or saprophytic species were identified. Pathogenic root fungi included Fusarium spp. (40% of all isolations), Macrophomina, Pythium, Rhizoctonia, and Sclerotium. Fusarium and Rhizoctonia were the most common fungal species identified and included F. oxysporum, F. verticillioides and F. solani, the three Fusarium species pathogenic on cotton plants. Population densities of Fusarium were not significantly different among locations or tissue types sampled. Macrophomina was isolated at greater numbers near the end of the growing seasons. Anastomosis groups of R. solani isolated from roots and soil debris included AG-3, -4, -7, 2-2, and -13 and anastomosis groups of binucleate Rhizoctonia included CAG-2, -3, and -5. Occurrences and frequency of isolations among sampling dates were not consistent. Fluctuations in the frequency of isolation of Rhizoctonia did not correspond with changes in frequency of isolation of the biological control fungus, Trichoderma. When individual or pooled frequencies of the mycobiota were compared to nematicide treatments, no specific trends occurred between treatments, application methods or rates. Results from this study show that use of 1,3-D and aldicarb in cotton fields does not significantly impact plant pathogenic fungi or saprophytic fungal populations. Thus cotton producers need not adjust seedling disease control measures when these two nematicides are used.

A 2-year (1999-2000) study was conducted at Starkville and Stoneville, MS to determine if the occurrence of the mycoflora varied on Roundup Ready (transgenic) compared to conventional soybean (Glycine max) cultivars. A total of 7,658 fungal isolates were identified from the pod and seed tissues of four cultivars compared at growth stages R6 and R8. Ninety-nine percent of all fungi isolated were mitosporic fungi and ascomycetes. In both years, total fungal isolates from the two locations were greater from the pod (65%) than from seed (33%) tissues. Isolation frequency from conventional cultivars was 54% compared to 46% for the transgenic cultivars. The most common fungi identified that are reported pathogens of soybean included Alternaria, Cercospora, Cladosporium, Diaporthe, Fusarium and Verticillium spp. When main effects and interactions were compared among the frequency data for the fungal genera, significant differences occurred, but consistent trends were not noted. Isolation frequencies of Diaporthe spp. during the R6 growth stage, were significantly greater on the conventional than on the transgenic cultivars in both years of the study, but only at Starkville. Isolation frequencies from samples taken during the R8 growth stage were similar at both locations in 1999 and 2000. Fusarium spp. isolated at R6 and R8 growth stages from pod and seed tissues were significantly greater on conventional than on transgenic cultivars in 2000. Even though frequencies were often significantly different between the transgenic and conventional cultivars, the data was not consistent between locations, pod and seed tissues, or growth stages. The pod and seed mycoflora of transgenic and conventional soybean cultivars was, therefore, similar in Mississippi.

Survival of the charcoal rot pathogen (Macrophomina phaseolina) in soybean (Glycine max) on residual root systems was studied over a 2-year period. Root segments colonized by M. phaseolina were placed into fiberglass-mesh bags and buried at depths of 0, 7.6, and 25.4 cm in a Marietta fine sandy loam soil in field microplots. Samples for year 1 and year 2 were buried in October 1999 and 2000, respectively, and sampled every 2 months over a 14-month period. Mean percent frequencies from both years for M. phaseolina showed a decreased linear trend over time from 91% before burial to 24.7% 2 months later, 10.3% at 6 months, and 0.3% after 14 months. Tissues were degraded in the June samplings of both years and this degradation was believed to be partially responsible for the reduced survival of M. phaseolina. Mean isolation frequencies were significantly (P ≤ 0.05) greater at the 0-cm depth than at 7.6 and 25.4 cm across all sampling dates. Other fungi which were present in the preliminary assay, including Fusarium spp. and those in the Phomopsis/Diaporthe complex, also declined by the end of the study. Trichoderma spp. was isolated at significantly (P ≤ 0.05) greater frequencies and a significant positive linear trend over time. It is possible that Trichoderma spp. were involved in direct tissue degradation and nutrient depletion of the root segments, or may have acted as a mycoparasite, reducing the survival of the pathogen M. phaseolina and other associated fungi. Results from this study indicate that farm practices which increase residue destruction immediately after harvest or those that enhance Trichoderma spp. populations may directly or indirectly lower the relative longevity of soilborne pathogens, including M. phaseolina.

ABSTRACT Rhizoctonia solani anastomosis group (AG)-13 was collected from diseased roots of field grown cotton plants in Georgia in the United States. Isolates of AG-13 did not anastomose with tester isolates of AG-1 through AG-12. Mycelium of all isolates of AG-13 were light brown but darkened as cultures aged. All isolates produced aerial mycelium. Concentric rings were visible after 3 to 4 days of growth but disappeared as cultures aged and darkened. Individual sclerotia were up to 1.5 mm in diameter, similar in color to the mycelium, and generally embedded in the agar. Clumps of sclerotia up to 5 mm in diameter were produced on the agar surface. All attempts to induce basidiospore production were unsuccessful. The 5.8S region of the rDNA from isolates of AG-13 was identical in length and sequence to isolates of all other AGs of R. solani. Length and sequence of the internal transcribed spacer (ITS) regions of rDNA from isolates of AG-13 were unique among AGs of R. solani. Similarity between AG-13 and other AGs of R. solani ranged from 68 to 85% for ITS region 1 and 85 to 95% for ITS region 2. Selected isolates of AG-13 caused minor or no damage to barley, cauliflower, cotton, lettuce, potato, and radish in laboratory or greenhouse studies.

A review of the fungi associated with soybean seeds, pods, or flowers was conducted in North America. Species of Deuteromycetes are the most common fungi in each of the soybean flower organs followed by the Ascomycetes and Phycomycetes which comprise about one-fourth of the total mycoflora. Eighty genera and about 135 or more species occur in seeds, pods, or flowers. With regard to numbers of taxa from separate mycofloras, 63 genera and about 108 or more species occur in seeds, 65 genera and about 88 or more species occur in pods, and 36 genera and approximately 47 or more species occur in flowers. Most of the fungi which occur in flowers can be cultured from pods, and the majority of those fungi occur in seeds. Methods for and a key are provided for the identification of the 30 most important selected fungi.

The efficacy of rye (Secale cereale) and wheat (Triticum aestivum) winter cover crops and cotton stalk and root destruction (i.e., pulling them up) were evaluated in field tests during two growing seasons for Hoplolaimus columbus management in cotton. The effect of removing debris from the field following root destruction also was evaluated. Wheat and rye produced similar amounts of biomass, and both crops produced more biomass (P

Fatty acid methyl esters (FAMEs) of isolates of Rhizoctonia solani AG-4 and AG-7 were characterized by gas chromatography and analyzed with Microbial Identification System software. Palmitic, stearic, and oleic acids were common in all isolates from both anastomosis groups (AGs) and accounted for 95% of the C14 to C18 fatty acids present. Oleic acid, most common in both R. solani AG-4 and AG-7 isolates, accounted for the greatest percentages of total FAMEs. The presence, quantities, or absence of individual fatty acids could not be used for distinguishing AG-4 and AG-7 isolates. Anteisopentadecanoic and 9-heptadecanoic acids, however, were specific to all three AG-7 isolates from Japan but absent in other AG-7 isolates and all AG-4 isolates. Pentadecanoic acid occurred in only two of the R. solani AG-4 isolates, but was not found in any of the AG-7 isolates. The AG-4 isolates could be distinguished from AG-7 isolates when quantities of FAMEs and key FAME ratios were analyzed with cluster analysis and principle components were plotted. Isolates of AG-7 from Arkansas, Indiana, and Georgia appeared to be more closely related to each other than to AG-7 isolates from Japan and Mexico. These differences in FAMEs were sufficiently distinct that isolate geographical variability could be determined. A dendrogram analysis cluster constructed from the FAMEs data showed results similar to that of the principal component analysis. Euclidean distances of total AG-4 isolates were distinct from total AG-7 isolates. The Arkansas and Indiana AG-7 isolates had a similar Euclidean distance to each another but the percentages were different for the AG-7 isolates from Japan and Mexico. In conclusion, variability of the FAMEs identified in this study would not be suitable as the main diagnostic tool for distinguishing individual isolates of R. solaniAG-4 from AG-7.

In a field study evaluating the diversity and density of the soilborne mycobiota in a cotton (Gossypium hirsutum L.) production system, Macrophomina phaseolina (Tassi) Goidanich was isolated on potato dextrose agar from dark brown to black lesions on feeder and secondary roots. Multiple proliferations of feeder and secondary roots were also observed. Isolate RB 656 obtained from these lesions was tested for pathogenicity in the greenhouse by mixing 25 ml of 2-week-old cornmeal sand inoculum (3 g of cornmeal, 100 g of sand, and 20 ml of distilled water) with 5 liters of autoclaved soil (Leefield loamy sand, pH 6.2) per pot (40 × 200 cm). Control pots containing autoclaved soil alone were included for comparison. On 17 September, 10 cotton seeds of DPL 90 were sown per pot. Each treatment had five replications. Forty days after planting, plant heights from pots containing M. phaseolina were lower (14.8 cm) than those in the control pots (19.6 cm), but stand counts were similiar. On this date, four plants were left in each pot to allow the remaining seedlings to reach full maturity. On 9 December, the four plants per replicate pot were removed and roots evaluated for damage. Lesions similiar to those seen originally were observed on the secondary and feeder roots of the infested pots, and the characteristic proliferation of feeder and secondary roots was noted. Tap roots in pots containing M. phaseolina were smaller (11.6 cm) than in the control pots (18.6 cm). Isolate RB 656 was reisolated from the damaged roots in the treated pots. This is the first report of M. phaseolina on cotton in Georgia.

During a study to determine the pathogenic fungi overwintering on dead cotton (Gossypium hirsutum L.) roots, two isolates of Rhizoctonia solani Kühn anastomosis group 7 (AG-7) were identified. Isolate #213 was obtained from dead roots near Tifton, GA, and isolate #219 was cultured from cotton roots near Midville, GA. Rhizoctonia solani AG-7 was previously reported in Arkansas, Indiana, and Asia (1). Isolates #213 and #219 were tested in the greenhouse for pathogenicity by mixing 25 ml of 2-week-old cornmeal sand inoculum (3 g of cornmeal, 100 g of sand, and 20 ml of distilled water) into 20 × 100 cm pots containing 2.25 liters of sterile soil (Tifton loamy sand, pH 6.1) per pot. Pots with noninfested soil were included as a control. Eight seeds of cotton (Delta and Pineland 90 DPL 90) were sown per pot. Each pot was a replicate and each treatment was replicated five times. At 20 days after planting, plant stands in soil infested with isolate #213 or #219 averaged 2 to 3 or 4 to 5 plants per pot, respectively, while stands in noninfested soil averaged 7 to 8 plants per pot. Brownish colored, sunken lesions were observed on roots, hypocotyles, and cotyledons of plants from pots infested with R. solani AG-7. Isolates #213 and #219 were reisolated from plants grown in their respective treatments. This is the first report of R. solani AG-7 in Georgia. Reference: (1) R. E. Baird et al. Plant Dis. 80:1421, 1996.

In September, 1996, cotton bolls of NuCotn 35B, DPL 90, Hart 404, NuCotn 33B, and DPL 5415 grown in three eastern Georgia counties were found to contain internal lint rot within single or multiple locules. No external boll damage was observed. Damage was always more severe on NuCotn 35B. Internal lint damage occurred in bolls tips that formed abnormal fissures or openings nearest the sutures. Such bolls often contained an additional locule. In one field of NuCotn 35B, over 30% of the bolls containing the abnormal tip morphology had locular lint rot. The abnormal boll morphology described previously as supernumerary carpel syndrome (SCS) has been associated with internal lint rot (2). Rotted lint within the locules was usually reddish brown mixed with yellow and olive to olive-brown. Gram-negative bacteria were isolated from infected bolls on nutrient agar and shown to be Pantoea agglomerans on the basis of rod shape, yellow pigmentation, and utilization of glucose both in an oxidative and fermentative manner without gas production. These characteristics are typical of bacterial strains belonging to the Enterobacteriaceae (facultative anaerobes). Even though bacterial taxonomists are uncertain about the species identification of certain genera within this family, strains 92-52D and 92-52E, isolated from NuCotn 35B, were identified within the complex P. agglomerans (Ewing and Fife) Gavini, Mergaert, Beji, Mielcarek, Izard, Kersters, and De Ley (=Enterobacter agglomerans;=Erwinia herbicola (1). Previously, P. agglomerans was reported to cause lint rot of cotton locules associated with SCS and stinkbug (Euschistus impictiventris) injury (2). In California, the lint rot bacterium was isolated from the stylar canal, especially when SCS was present. To confirm pathogenicity of the isolates, 30 bolls of NuCotn 35B were excised, brought into the laboratory, and surface disinfested with 70% ethanol. Bolls were inoculated by dipping sterile dissection needles in bacterial colonies from 24-h cultures on nutrient agar and immediately puncturing (3- to 7-mm depth) 10 bolls per strain. An additional 10 bolls were wounded with sterile needles for comparison. The bolls were placed in a moist chamber and incubated at 28°C for 72 h. Locule damage was rated on a 0 to 3 scale in which 0 = no damage, 1 = trace damage, 2 = moderate discoloration and deterioration, and 3 = total discoloration and deterioration of the fiber. Bolls inoculated with strains 96-52D and 96-52E had mean boll rot indices of 2.0 and 1.5, respectively, and Koch's postulates were completed for both strains. The noninoculated bolls had a discoloration indice of 0.6, but 7 out of 10 were rated as 0. The infections of the three bolls by P. agglomerans may have been caused by natural inoculum that survived the surface disinfestation. These findings are the first report of P. agglomerans causing lint rot of bolls with SCS in Georgia. References: (1) J. Mergaert et al. Curr. Microbiol. 8:327, 1985. (2) G. M. Watkins, ed. Compendium of Cotton Diseases. American Phytopathological Society, St. Paul, MN, 1981.

The survival of the mycobiota on pod and stem debris of soybean produced in a no-tillage system with cover crops of alfalfa, canola, rye, or wheat or with no cover was studied during 1994 and 1995. Fiberglass mesh bags containing pods and stems were assayed every 28 to 31 days to determine the isolation frequency of fungi. Over 90% of the 11,906 isolates obtained were members of the Deuteromycotina. The most common genera isolated were Alternaria, Cercos-pora, Colletotrichum, Epicoccum, Fusarium, and Phoma. Alternaria spp. had the greatest isolation frequencies and constituted 40% of the total cultures. Numbers of total fungi (all fungi isolated) on sampling dates in 1994 were similar to the totals in 1995. In May 1994, the mean isolation rates for many of the fungal species were significantly lower (P = 0.05) in several of the cover crops, but no consistent pattern could be determined. Common soybean pathogens isolated included Colletotrichum spp., Diaporthe spp., and Cercospora kikuchii. Fusarium graminearum, which is responsible for several diseases of maize and wheat, was commonly isolated during this study. Of the Diaporthe spp. (anamorph Phomopsis spp.), 87% were identified as D. phaseolorum var. sojae. Colletotrichum spp. were identified as C. truncatum in 85% of the isolates, C. destructivum (teleomorph Glomerella glycines) in 12%, and both species in 3%. Cercospora kikuchii was more commonly isolated from pods than from stem tissue, and Colletotrichum spp. occurred more frequently on stems. Isolation frequencies of Diaporthe spp. were greater in May of both years than in the preceding months. These results show that no-tillage soybean debris harbors numerous fungi pathogenic to soybean, and producers who grow soybeans continuously may find more disease in this crop and lower yields. Fungi that attack crops such as maize and wheat were commonly isolated from soybean debris in both years, and a no-tillage rotation which includes maize or wheat could result in increased disease in these crops. Isolation frequencies of the fungi from cover crops varied with the sampling date, but no consistent patterns could be determined for a particular cover crop or fungal species. This is the first detailed study of survival rates of soybean, maize, and wheat pathogens that overwinter on soybean debris in a no-tillage system.

A survey was conducted to examine the geographical distribution of plant-parasitic nematodes in Georgia cotton fields. A total of 778 fields in 11 Georgia counties were sampled from 1 September through 15 December 1995. Four nematode genera parasitic on cotton were found in this survey: Meloidogyne spp., Rotylenchulus sp., Hoplolaimus sp., and Belonolaimus sp. Meloidogyne spp. was present in 9% to 56% of the fields in individual counties. Rotylenchulus sp. was found in 10 counties, Hoplolaimus sp. was found in 6 counties, and Belonolaimus sp. was found in 2 counties. From all of the samples collected for this survey, Meloidogyne spp. were found in 31% of the samples, Rotylenchulus sp. was found in 14%, Hoplolaimus sp. was found in 7%, and Belonolaimus sp. was found in 0.3%. Burke County had the greatest number of fields infested by at least one of these genera (67%) and the greatest number of fields above Georgia's action thresholds (38%). Laurens County had the fewest fields where these genera were present (13%), and only 3% of fields had nematode populations above threshold levels. Data from samples collected from cotton fields and submitted by county agents from 1993 through 1994 were compiled to provide historical information about nematode distribution and population density. The results from this survey show that the major nematodes damaging to cotton are not present in all counties in Georgia. Counties in which cotton has historically been a major crop are likely to have higher levels of Meloidogyne spp., Hoplolaimus sp., and Rotylenchulus sp. in current cotton crops. Counties in which soybean has historically been a major crop are likely to have higher levels of Hoplolaimus sp. and Rotylenchulus sp. in current cotton crops.

Sixteen species of fungi were isolated from the pericarp, endosperm, scutellum, and embryo of low (<75% germination), moderate (75-89%), and high vigor (>89%) seedlots of supersweet sweet corn (sh2) hybrids over two years. Most frequently isolated species werePenicillium oxalicum, Rhizopus arrhizus (14.5%), andRhizopus spp (17.4%).Fusarium moniliforme, a common inhabitant in field or dent corn, was isolated from only 2.4% of the samples. The low mean isolation values may be the result of poor conditions for infection or the data may reflect the methods used including sterilization techniques or random tissue selection.Aspergillus niger, F. moniliforme, andP. oxalicum, were isolated at a significantly greater level than other fungi from the high vigor hybrids at 0.89, 0.46 m and 4.46 respectively compared to 0.04, 0, and 1.82 for the low vigor hybrids. When Total Fungi were compared mean isolations were significantly greater from the high vigor hybrids at 11.96, the moderate 8.79, or low 4.86 vigor groups. When data from seed sources for all vigor groups were compared, significantly greater mean isolations were obtained from Illinois Foundation Seed hybrids forCladosporium sp,R. arrhizus, andRhizopus spp., but greater rates were obtained forFusarium oxysporum from the Asgrow hybrids. Isolation frequencies for the 16 species were not significantly different between the seed tissue types from any of the hybrids evaluated during this investigation. Results from this study showed that there is a diverse group of fungi present within thesh2 seed and seed treatments must be developed which will minimize seed rot and seedling blight from both internal seedborne and external pathogens.

Twenty-five female, 23-month-old Long-Evans rats were sacrificed for investigation of correlations between serum prolactin levels, pituitary weights, numbers and ultrastructural features of pituitary lactotrophs. Serum prolactin levels measured by radioimmunoassay varied from 7.1 ng/ml to 1,534.0 ng/ml. Pituitary weights ranged from 16.0 mg to 255.0 mg. Serum prolactin levels were increased when pituitary weights were high; statistical analysis revealed a correlation between the two. No significant correlation was evident between serum prolactin levels and lactotroph percentage and ultrastructure. Light microscopy revealed a large number of lactotrophs in almost every pituitary. Values for nuclear, cell and cytoplasmic areas, nuclear/cytoplasmic ratios and nucleolar diameters remained similar in the lactotrophs of all animals. Electron microscopy disclosed no lactotroph damage or vascular injury. Ultrastructural morphometry of lactotrophs in the pituitaries studied showed no major differences in cytoplasmic volume densities of endoplasmic reticulum, Golgi complex, mitochondria, secretory granules and lysosomes as well as secretory granule diameters. It is reasonable to conclude that elevations of serum prolactin levels are due to an increase in pituitary mass, resulting from accumulation of lactotrophs. Thus hyperprolactinemia appears to be secondary to an increase in the number of lactotrophs and not hyperactivity of individual cells.

Review of this case resulted from the realization of potential pitfalls of field and dispensary care in evaluating and treating penetrating chest trauma (PCT) at high altitude. Literature review of PCT pertaining to this case is followed by a discussion of proper and improper field care, medical evacuation, stabilization, and disposition for service members who sustain PCT. The Mountain Warfare Training Center Tension Pneumothorax Potential Scale is introduced. Recommendations for appropriate primary care of victims of PCT, based on the discussion, are made.