Promising primers for detection of phytoplasma causing coconut lethal yellowing disease in Mozambique

Coconut is one of the main cash crop in Mozambique, which occupied the second position after Tanzania in coconut production in Africa. Coconut production was drastically affected by the occurrence of a devastating Coconut Lethal Yellowing Disease (CLYD) epidemics, which reduced significantly the coconut yields. CLYD symptoms triggered upon phytoplasma infection, i.e. premature fruit dropping, necrosis of the inflorescence and progressive yellowing of the leaves, are used to identify infected trees. However, the diagnostic based uniquely on symptoms is not conclusive to confirm infection, and needs to be confirmed by molecular methods. In this study, three previously described reference primers for phytoplasma detection were tested on infected samples collected in Mozambique. Since those primers gave incongruent results, 20 new primer pairs targeting the 16S rDNA region, were newly designed. To evaluate their performance in detecting coconut infecting phytoplasma, 108 samples were tested and selected positive samples confirmed by sequencing. Our results showed a new primer pair more accurate and reliable compared to the reference pairs for CLYD detection in Mozambique. Moreover, the new primer pair was able to detect a new putative phytoplasma variant in Mozambique. Therefore, this study makes an important contribution to CLYD phytoplasma molecular diagnostics and its causative agent, giving insights that may be applied to the study of CLYD phytoplasma infection systems.     

Two cases of cutaneous nodular dirofilariasis in the cat

Cutaneous nodular dirofilariasis is occasionally reported in dogs but is rare in cats. This report describes two cases of cutaneous nodular lesions due to Dirofilaria immitis adult parasites in domestic shorthaired cats living in an endemic area in northern Italy.     

Root hydraulic conductivity and root growth capacity of black spruce (Picea mariana) seedlings

The relationship between root elongation and root hydraulic conductivity was investigated in 1-year-old, overwintered black spruce (Picea mariana (Mill.) seedlings. Hydraulic conductivity was estimated by observing water flux through decapitated roots under positive pressure. Five hydraulic conductivity parameters were estimated: (1) water flux under a minimal pressure of 0.2 MPa (J(v, min)); (2) pressure at which a linear relationship between water flux and pressure began (P(min)); (3) slope of the linear water flux-pressure relationship (L(v)); (4) pressure at which the linear relationship between water flux and pressure ended (P(max)); and (5) maximum water flux (J(v, max)). Between day 1 and day 2 after thawing of the growing medium, there were significant increases in L(v) and J(v, max) but there was little, if any, root elongation. Root elongation averaged 5.4 cm seedling(-1) 5 days after thawing and 88.2 cm seedling(-1) 20 days after thawing. Root hydraulic conductivity increased with new root length initially. But between days 20 and 30, when new root length nearly doubled, P(min) was the only measure of hydraulic conductivity that increased significantly. There were significant correlations (P < 0.05) between white root length and both J(v, min) (r(2) = 0.90) and J(v, max) (r(2) = 0.91).     

An overview of Ontario's Stock Quality Assessment Program

Since 1992, the Stock Quality Assessment Program at the Ontario Forest Research Institute (OFRI) has offered seedling physiological testing services to foresters and nursery managers. The purpose of this program is to improve nursery stock quality and plantation performance by developing and applying procedures for assessing the physiological quality of nursery stock. Two levels of testing are available: Seedling Certification and Problem Stock Testing. Testing at both levels involves a visual assessment, measurement of chlorophyll fluorescence of the foliage and root growth potential. Applying these tests has directly improved plantation establishment between 1992 and 1995 by preventing over 3 million damaged seedlings from being planted, at an estimated regeneration cost savings of over $2 million (Can). Consideration of cost-benefits for both direct and indirect values demonstrates the merits of a stock testing program. Testing has been conducted either at OFRI or at a private lab, providing clients with an impartial assessment of their stock. Consistent test results, comparable from year-to-year and between laboratories, are achieved by the use of controlled environment testing, trained personnel and duplicate testing on selected stocklots. A database comprised of physiological test information for over 1100 stocklots provides a basis for comparing and ranking seedlings grown throughout the province. This database may be used to refine operational nursery practices, to evaluate changes in seedling quality over time, and to relate seedling quality to field performance.     

Acquired thermotolerance of jack pine, white spruce and black spruce seedlings

The acquired thermotolerance of first-year seedlings of jack pine (Pinus banksiana Lamb.) hardened at 36, 38, 40 or 42 degrees C for 90, 180 or 360 minutes and of black spruce (Picea mariana (Mill.) B.S.P.) hardened at 34, 36, 38 or 40 degrees C for 30, 90, 180 or 360 minutes was determined by comparison of needle damage to that of non-hardened seedlings (25 degrees C) following exposure to temperatures of 49 and 47.5 degrees C, respectively. Compared to seedlings kept at 25 degrees C, heat injury sustained from exposure to high temperatures was markedly reduced following hardening for 180 minutes at 36 and 38 degrees C in jack pine and black spruce, respectively. Increasing the exposure time at 36 degrees C in jack pine, and at 36 to 40 degrees C in black spruce, also reduced needle damage. The duration of increased thermotolerance was investigated in jack pine, black spruce and white spruce (Picea glauca (Moench) Voss) by comparing heat injury from high temperatures in non-hardened seedlings and in seedlings hardened at 38 degrees C for 180 minutes a day for either 1, 3 or 6 days. In all three species, the duration of acquired thermotolerance increased with the number of days of heat hardening. For jack pine and white spruce seedlings hardened at 38 degrees C for 6 days, increased thermotolerance persisted for at least 14 and 10 days, respectively, after the end of the hardening treatment. In contrast, the thermotolerance of black spruce seedlings hardened at 38 degrees C for 6 days remained elevated for only 4 days.     

Papular eosinophilic/mastocytic dermatitis (feline urticaria pigmentosa) in Devon Rex cats: A distinct disease entity or a histopathological reaction pattern?

A maculopapular eruption with clinical and histological features similar to those previously described in Sphinx cats under the name of urticaria pigmentosa is reported in five unrelated Devon Rex cats. Physical examination revealed erythematous, occasionally crusted papules, with a bilaterally symmetrical linear distribution on the latero-ventral trunk in two cases and a diffuse distribution on the ventral thorax in the other three cats. One cat also had a greasy seborrhoea on the head and dorsum. Pruritus and pigmented macules were present only in cats affected by secondary bacterial infection. Histological examination of papules in all cats and of the lesional skin of the cat affected by greasy seborrhoea revealed the presence of a perivascular to diffuse mastocytic and eosinophilic infiltrate in the dermis. The mean numbers of nondegranulated and degranulated mast cells per mm(2) were 303.2 and 451.6, respectively. The condition waxed and waned in all cats, and exacerbations were controlled with prednisolone or essential fatty acids.     

A Preliminary Trial Using Multi-target Polymerase Chain Reaction (multiplex PCR) and Restriction Fragment Length Polymorphism (PCR-RFLP) on the Same Feedstuffs to Detect Tissues of Animal Origin

A preliminary study using multi-target polymerase chain reaction (multiplex PCR) and restriction fragment length polymorphism (PCR-RFLP) was done on the same feedstuffs to detect animal tissues. The results of the two methods differ somewhat: PCR-RFLP did not detect any signal in any sample, but multiplex PCR detected a signal in one sample. These findings could be a basis for further investigations.     

Morphological indices for canine spermatozoa based on the World Health Organization laboratory manual for human semen

Attempting to contribute to the development of a more objective morphological evaluation of dog spermatozoa, in this study the indices of multiple sperm defects (multiple abnormalities index [MAI]; teratozoospermic index [TZI]; sperm deformity index [SDI]) were calculated following the World Health Organization (WHO) guidelines. In Experiment I, the concordance of MAI, TZI and SDI with the proportions of morphologically normal spermatozoa (MNS) was evaluated in fresh ejaculated spermatozoa (dogs = 47). In Experiment II, the potential role of indices as prognostic values was assessed in spermatozoa of different origin and treatment (fresh ejaculated: n = 6; fresh epididymal: n = 6; frozen‐thawed ejaculated: n = 6) by their correlation with different semen parameters (motility, membrane integrity and acrosome status) and with an in vitro sperm function test. Samples with different proportions of MNS showed different values of SDI, the index that better represented the decline of sperm morphology in both fresh and frozen‐thawed samples (Exp. I and II; p < 0.05). No correlations between indices and semen parameters were observed (Exp. II), but when samples were evaluated collectively, negative correlations (SDI‐motility, p = 0.01; SDI‐acrosome integrity, p = 0.002) were found. Including all the defects of each spermatozoon, SDI might be a useful index during morphological analysis and better discriminates the increase in multiple defects. A more objective morphological evaluation for dog spermatozoa was achieved by the WHO method, and in vitro tests allowed to elucidate the validity of SDI as prognostic indicator of in vitro fertilizing potential.