New Mexico and the southwestern US are affected by a unique population of tomato spotted wilt virus (TSWV) strains.

Tomato spotted wilt virus (TSWV) is an important pathogen of many ornamental, greenhouse and agronomic crops worldwide. TSWV also causes sporadic problems in a number of crops in New Mexico (NM). Nucleocapsid gene sequences obtained from six different crop species across the state over four different years were used to characterize the NM TSWV population. This analysis shows that NM is affected by a unique TSWV population that is part of larger independent population present in the southwestern US. This population likely arose due to geographic isolation and is related to other TSWV populations from the US, Spain, and Italy.

First Report of Anthracnose of Sunflower Sprouts Caused by Colletotrichum acutatum in New Mexico.

In December 2011, edible sunflower sprouts (Helianthus annus) of two different commercially grown cultivars (Sungrown and Tiensvold) exhibiting stem and cotyledon lesions were submitted to the New Mexico State University Plant Clinic for disease diagnosis. The sample originated from an organic farm in Santa Fe County where the grower utilizes a small indoor growing facility. Stem lesions were elongate, reddish brown, and often constricted, resulting in stem girdling. Lesions on the cotyledons were dark brown with tan centers and round to irregular in shape. In some cases, the entire cotyledon was blighted. Fungal hyphae were observed on some lesions using a dissecting microscope. Colletotrichum acutatum was isolated from stem and cotyledon lesions when symptomatic tissue was plated on water agar. Conidia were fusiform ranging from 6.4 to 18.4 µm long and 2.1 to 5.1 µm wide and averaged 11.9 µm × 3.4 µm. Spores were measured from cream-colored colonies produced on acidified potato dextrose agar. PCR amplification and sequence analysis of 5.8S ribosomal DNA and internal transcribed spacers I and II was performed using primers ITS4 and ITS6 (2). An amplification product of approximately 600 base pairs in size was directly sequenced (GenBank Accession No. JX444690). A BLAST search of the NCBI total nucleotide collection revealed a 99% identity to multiple C. acutatum (syn: C. simmondsii) isolates. Four isolates were identified as C. acutatum based on morphological characteristics and DNA analysis. Koch's postulates were performed using four isolates of the pathogen and the two commercial sunflower cultivars (Sungrown and Tiensvold) originally submitted for disease analysis. Sunflower seeds were imbibed in distilled water for 24 h then sewn into peat plugs. Prior to seed germination, 5 ml of a C. acutatum spore solution (1 × 106/ml) from each isolate was applied to five peat plugs using an atomizer. Control plants were inoculated with distilled water and otherwise treated identically. Both sunflower cultivars were inoculated with each isolate of the pathogen and the test was replicated twice. The sewn peat plugs were incubated for 5 days at 21°C and 50% relative humidity. Symptoms similar to the original samples were present on 100% of the sprouts after 5 days. PCR and sequence analysis performed on cultures obtained from lesions showed a 100% match to the original New Mexico isolates fulfilling Koch's postulates. In an indoor organic facility, such as the one in NM, this disease has the potential to be very difficult to manage and the potential to infect a high percentage of the crop resulting in significant economic losses. To our knowledge, this is the second report of C. acutatum on sunflower sprouts in the United States (1) and the first report in New Mexico. References: (1) S. T. Koike et al. Plant Dis. 93:1351, 2009. (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, San Diego, 1990.

First Report of Phytophthora nicotianae on Bulb Onion in the United States.

Phytophthora nicotianae (synonym P. parasitica) Breda de Haan was isolated from recently harvested onion bulbs (Allium cepa) in cold storage from a commercial field in southern New Mexico. Deteriorating, water-soaked tissue from the center of four bulbs was plated onto water agar and incubated at room temperature. After 72 h, cultures of Phytophthora (identified by the presence of coenocytic hyphae and papillate sporangia) were isolated and transferred to V8 agar amended with ampicillin (250 mg/liter), rifampicin (10 mg/liter), and pimaricin (0.2% wt/vol). Isolates were identified as P. nicotianae based on morphological characteristics and DNA analysis. Sporangia were sharply papilliate, noncaducous, and ovoid to spherical. The average sporangium size was 45.9 × 39.9 µm with a length-to-width ratio of 1.15. Clamydospores, both terminal and intercalary, were spherical to ovoid and averaged 37.2 × 35.2 µm (2). PCR from whole-cell extracts was performed on four cultured isolates from the infected onion tissue using previously described primers ITS4 and ITS6, which amplify the 5.8S rDNA and ITS1 and ITS2 internal transcribed spacers (1,4). A band of approximately 890 bp was amplified and directly sequenced (GenBank Accession No. HQ398876). A BLAST search of the NCBI total nucleotide collection revealed a 100% similarity to multiple P. nicotianae isolates previously sequenced (1). To confirm the pathogenicity of the isolates, onion seedlings were inoculated with 25 ml of P. nicotionae zoospore solution (15,000 zoospores/ml). Necrosis of leaf tissue and seedling death was observed 5 days postinoculation. P. nicotianae was reisolated from the infected onion seedlings and the ITS region was sequenced to confirm its identity. P. nicotianae was previously reported in bulb onion from Australia, Taiwan (Formosa), and Zimbabwe (Rhodesia) (2). P. nicotianae was reported on bunching onions (A. fistulosum) in Hawaii in 1989 (3). Onions are an important crop in New Mexico with a total production value of 47 million dollars in 2008 (NM Agriculture Statistics 2008). This discovery of a potentially significant postharvest disease poses a threat to the onion industry in New Mexico. To our knowledge, this is the first report of P. nicotianae in bulb onion in the United States and the first report of P. nicotianae in New Mexico on any crop. References: (1) D. E. L. Cooke and J. M. Duncan. Mycol. Res. 101:667, 1997. (2) D. C. Erwin and O. K. Ribeiro. Page 56 in: Phytophthora Diseases Worldwide. The American Phytopathological Society, St Paul, MN, 1996. (3) R. D. Raabe et al. Information Text Series No. 22. University of Hawaii. Hawaii Inst. Trop. Agric. Human Resources, 1981. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, 1990.

First Report of Buckeye Rot Caused by Phytophthora nicotianae in Tomato in New Mexico.

Phytophthora nicotianae Breda de Haan was isolated from turning tomato fruit (Solanum lycopersicum L.) in August 2010 from a garden in central New Mexico. Symptoms typical of buckeye rot including brown, water-soaked, necrotic lesions with concentric rings were observed on three tomato fruit. Tissue from each fruit was surface sterilized and plated onto water agar and incubated at room temperature. After 72 h, colonies of Phytophthora (identified by the presence of coenocytic hyphae and papillate sporangia) were found and subcultured by hyphal tips to V8 agar amended with ampicillin (250 mg/liter), rifampicin (10 mg/liter), and pimaricin (0.2% wt/vol). The isolates of Phytophthora were identified as P. nicotianae based on morphological characteristics and DNA analysis. Sporangia were sharply papillate, noncaducous, and ovoid to spherical. The average sporangium size was 44.5 × 35.5 µm with a length-to-width ratio of 1.26. Chlamydospores, both terminal and intercalary, were spherical to ovoid and averaged 38.9 × 37.5 µm. PCR amplification and sequence analysis on three isolates from the infected tomato tissue was performed using primers ITS4 and ITS6 that amplify the 5.8S rDNA and ITSI and ITSII internal transcribed spacers (1,2). A band of approximately 890 bp was amplified and directly sequenced (GenBank Accession No. HQ711620). A BLAST search of the NCBI total nucleotide collection revealed a 100% similarity to multiple P. nicotianae isolates previously sequenced. Pathogenicity tests with sequenced P. nicotianae isolates were performed to confirm virulence on tomato fruit. Tomatoes were surface sterilized with 95% ethanol and 0.1 ml of a P. nicotianae zoospore suspension (10,000 zoospores/ml) or sterile water was pipetted onto the surface of the tomato fruit. After 5 days in a humidity chamber, all three inoculated tomatoes had expanding water-soaked, circular lesions and the negative control showed no disease symptoms. P. nicotianae was successfully reisolated from the inoculated tomato tissue and the ITS region was sequenced to confirm its identity. Although the disease has been reported in many other states since the early 1900s, to our knowledge, this is the first report of P. nicotianae causing disease on tomato in New Mexico. References: (1) D. E. L. Cooke and J. M. Duncan. Mycol. Res. 101:667, 1997. (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, San Diego, 1990.

First Report of Septoria Leaf Spot of Pistachio in New Mexico.

In September of 2008, a Septoria sp., the causal agent of Septoria leaf spot of pistachio (Pistacia vera L.) was isolated from leaf lesions in an orchard in southern New Mexico. Tree fruit and nut crops including pistachios are becoming an increasingly important part of New Mexico's agricultural industry with total cash receipts of $103 million in 2007 (3). This preliminary positive for Septoria prompted a survey of pistachio-growing counties in the state. The surveyed orchards accounted for approximately 30% of the pistachio acreage in New Mexico. Results indicated that all five pistachio-growing counties had orchards infected with a Septoria sp. Isolates of Septoria from leaf lesions were identified as Septoria pistaciarum Caracc. based on the following symptoms and morphological characteristics of the fungus: leaf lesions were usually circular, 0.5 to 3 mm in diameter, and contained many pycnidia per lesion; pycnidia were dark, ostiolate, and measured 101 to 255 × 69 to 133 µm; and conidia were hyaline, filiform, contained 3 to 9 septa, and measured 3 to 4 × 60 to 149 µm. Most orchards were only mildly affected. In severe cases, hundreds of leaf lesions were present on diseased leaves; large sections of the leaves turned tan and some trees defoliated prematurely. This widespread occurrence of Septoria leaf spot in New Mexico in 2008 suggests that the disease had already been present in the state for several years. A higher average rainfall in the summer of 2008 provided excellent conditions for disease development. Because of the high amounts of inoculum currently present in New Mexico orchards, Septoria leaf spot may emerge as a recurring disease problem for pistachio producers. This disease was first reported in the United States in Texas in 1971 and was also reported in Arizona in 1989 (1,2,4). To our knowledge, this is the first report of Septoria leaf spot of pistachio in New Mexico. References: (1) A. Chitzandis. Ann. Inst. Phytopathol. Benaki 10:29, 1956. (2) J. L. Maas et al. Plant Dis. Rep. 55:72, 1971. (3) New Mexico Agricultural Statistics, Department of Agriculture, 2007. (4) D. J. Young and T. Michailides. Plant Dis. 73:775, 1989.

Xylella fastidiosa Detected in New Mexico in Chitalpa, a Common Landscape Ornamental Plant.

Different strains of Xylella fastidiosa cause a variety of significant disease problems in agricultural and ornamental plants, including Pierce's disease in grapes, oleander leaf scorch, pecan bacterial leaf scorch, and alfalfa dwarf disease. X. fastidiosa has never been reported in New Mexico but is known to exist in surrounding states (California, Arizona, and Texas). During the summer of 2006, several chitalpa (Chitalpa tashkinensis) hybrid trees with leaf scorch symptoms and branch die back were observed in Las Cruces, NM and they tested positive for X. fastidiosa by ELISA. Additional samples from these plants and others were analyzed by ELISA, PCR (2), and cultured on XfD2 medium (1). Known positive and negative oleander samples from Arizona were included as controls. Fifteen of thirty tested chitalpa were PCR and ELISA positive, indicating that they were infected with X. fastidiosa. Bacterial colonies that were PCR positive were also recovered from 10 of the XF positive samples that were plated. DNA sequences of PCR products amplified from chitalpa and isolated bacterial colonies (GenBank Accession Nos. EF109936 and EF109937) were identical to each other, 97% similar to X. fastidiosa strain JB-USNA, and 96% similar to the Temecula 1 strain. Independent ELISA testing (Barry Hill, California Department Food and Agriculture, Sacramento, CA) confirmed our ELISA and PCR results. On the basis of these results, we conclude that X. fastidiosa is present in New Mexico and that the common landscape ornamental chitalpa is a host for X. fastidiosa. Additional work is required to determine if X. fastidiosa is pathogenic to chitalpa and to examine the relevance of this potential X. fastidiosa reservoir to agricultural production in New Mexico and other areas where chitalpa is grown. References: (1) R. P. P. Almeida et al. Curr. Microbiol. 48:368, 2004. (2) M. R. Pooler et al. Lett. Appl. Microbiol. 25:123, 1997.

The development of ischemic heart disease in relation to autoimmune thyroid disease in a 20-year follow-up study of an English community.

The original Whickham Survey documented risk factors for cardiovascular disease and the prevalence of thyroid disorders in a sample of 2779 adults that closely matched the British population. A 20-year follow-up study has determined outcomes in terms of morbidity and mortality from ischemic heart disease in over 97% of the original survey population. Analysis of deaths from all causes and from ischemic heart disease showed no association with antithyroid antibody status identified at first survey. A multiple logistic regression using the development of ischemic heart disease in the total population at follow-up as the dependent variable found that the significant predictor variables for men were age, cholesterol, mean arterial blood pressure, smoking history, and skinfold thickness index. For women only age, cholesterol, and mean arterial blood pressure were significant. The presence of autoimmune thyroid disease, as defined by either hypothyroidism, positive antithyroid antibodies, or raised serum thyrotropin at first survey, was not significant. A retrospective cohort study of a subsample of women identified at first survey with positive antithyroid antibodies or raised serum thyrotropin and closely matched controls found no significant association with mortality or development of ischemic heart disease. There is no evidence from this study to suggest that evidence of autoimmune thyroid disease identified 20 years ago is associated with an increased risk of ischemic heart disease.

Effect of the risk factors for stroke on survival.

In a case-controlled study into the risk factors for admission to hospital with stroke, 400 subjects and 400 age and sex-matched controls were recruited. All bar two subjects were followed until death or 6 months. Previous stroke and regular snoring (p = 0.0013 and p less than 0.0001 respectively) were the only two risk factors adversely to effect mortality. Transient ischaemic attack, ischaemic heart disease, hypertension, atrial fibrillation, diabetes mellitus did not significantly effect prognosis. An apparent beneficial effect of drinking alcohol and smoking became insignificant when the confounding influence of age was taken into account.

Ectrodactyly and syndactyly in a common marmoset (Callithrix jacchus).

This paper describes previously unreported malformations of both fore and hind feet in a liveborn common marmoset. Ectrodactyly of both fore feet and left hind foot and syndactyly of the right hind foot were observed.

A pheromonal function for the perineal skin glands in the cow.

Two cows with synchronised oestrous cycles were housed with a Friesian bull. During the midcycle period of the cows, one cow was given an intradermal injection of adrenaline in the perineal region. This caused local sweat gland discharge. The other cow was treated with water. The number and types of bull-to-cow behaviours were recorded before, during and after the day of treatment. This was repeated at three-weekly intervals for a total of four times. On the day of treatment the bull directed a greater proportion of olfactory behaviours towards the adrenaline-treated cow than the other cow (P less than 0.05). There was no difference between the cows before or after the day of treatment. The above experiments were repeated with a Hereford bull who showed the same response as the Friesian for the first two but not the last two experiments. The increased proportion of bull olfactory behaviours elicited by an increased perineal skin gland discharge, adds support to our hypothesis that the perineal skin glands are the source of an oestrous pheromone in the cow.

The prognostic value of haematocrit in acute stroke.

One hundred and twenty-two consecutive patients admitted with acute stroke in a carotid distribution had venous blood taken for haematocrit (Hct), haemoglobin (Hb), white cell count (WCC) and urea estimations. Patients were followed for 12 weeks to determine the influence of haematocrit upon fatality. There were 96 patients aged greater than or equal to 65 years and 26 patients less than 65 years. No association could be demonstrated between Hct levels and fatality at 4 or 12 weeks. Regression analysis demonstrated that only increasing age (P less than 0.05) and a raised WCC (P less than 0.005) were independent factors significantly associated with fatality at both 4 and 12 weeks. In the elderly stroke patient (greater than or equal to 65 years) only WCC was significantly associated with fatality (P less than 0.005). Haematocrit levels are of no prognostic value for fatality in acute stroke. A raised white cell count is an important and independent prognostic factor for fatality at both 4 and 12 weeks following stroke.

Longitudinal study of central motor conduction time following stroke. 1. Natural history of central motor conduction.

A 12 month longitudinal study has been performed on 118 subjects following first-ever stroke to determine changes in central motor conduction time (CMCT) to upper limb muscles. The responses to electromagnetic stimulation of the motor cortex and cervical motor roots were recorded bilaterally in the surface electromyograms of pectoralis major, biceps and triceps brachii and thenar muscles. The CMCTs obtained from these recordings in stroke patients have been compared with those obtained in 53 normal healthy subjects of a similar age. The first measurements were made within the immediate post-stroke period (12-72 h of the onset of symptoms) and repeated at set time intervals over 12 months. The first assessment of CMCT identified three groups: those with normal responses, delayed responses and absent responses. During the first 12 months following stroke various changes in CMCT occurred. Central motor conduction time may remain unchanged, delayed CMCT may return to normal and previously absent responses may reappear and be delayed or normal. Using electromagnetic stimulation of the motor cortex the thresholds for motor evoked responses in the different muscles were initially high and fell over 12 months.

Longitudinal study of central motor conduction time following stroke. 2. Central motor conduction measured within 72 h after stroke as a predictor of functional outcome at 12 months.

A longitudinal study was performed on 118 first-ever stroke patients to evaluate neurophysiological measurements of central motor conduction time (CMCT) in the period immediately following stroke as predictors of functional outcome and mortality at 12 months. Measurements of CMCT were made as described in the accompanying article (Heald et al., 1993, Brain, 116, 1355-1370), in which the following three groups of patients were recognized within 12-72 h after the onset of symptoms: normal response group, delayed response group and no response group. Neurophysiological and clinical investigations were commenced 12-72 h (designated as day 1) after the onset of symptoms and repeated at set time intervals up to 12 months. The subjects were examined neurologically and assessed using the Motricity Index for muscle strength, the Nine-hole Peg Test to measure manual dexterity, the Barthel Score for activities of daily living and the modified Rankin Scale for functional outcome. The duration of stay in hospital and the occurrence of stroke-related death were noted. During the first week following stroke, absence of responses correlated closely with the patient's symptoms and neurological observations of abnormal muscle tone and tendon reflexes. Correlations were made in the three groups of patients of functional scores at day 1 and at 12 months. Patients with normal CMCT had consistently higher scores throughout the 12 month period and achieved significantly better functional recovery. Patients with no responses showed poor performance in neurological and functional tests throughout the 12 month period. Patients with delayed CMCT had neurological and functional scores intermediate between those of the other two groups, but outcome at 12 months was similar to those in the normal response group. Where the threshold to cortical stimulation was abnormally high, functional outcome was generally poor. Mortality was highest in the group with absent responses and the survivors spent the longest period in hospital. In conclusion, the observation of normal or delayed CMCT at day 1 identifies a group of patients with a high probability of survival and functional recovery. The absence of responses to cortical stimulation at day 1 identifies a group of patients who are at high risk of poor functional recovery at 12 months and greater probability of stroke-related death during this period.

Historical risk factors for stroke: a case control study.

The aim of this study was to determine the risk factors for admission to hospital with stroke by means of a case-control study recruiting age- and sex-matched controls from the local community. Four hundred pairs of patients and controls were recruited. Of historical factors, preceding cerebrovascular disease contributed the greatest odds of stroke (odds ratio 9.8). Taking prescribed medicines (odds ratio 2.6), regular snoring (odds ratio 3.2), smoking (odds ratio 1.7) and some factors in the family history were also significant risk factors.